Pharmacologically active 10alpha-methyl, 9beta-steroids of the pregnane series



United States Patent 3,373,172 PHARMACOLOGICALLY ACTIVE Illa-METHYL, 9,8- STEROIDS OF THE PREGNANE SERIES Engbert Harmen Reerink, Pieter Westerhof, and Hendrik Fredrik Louis Scholer, Van Houtenlaan, Weesp, Netherlands, assignors to North American Philips Company, Inc., New York, N.Y., a corporation of Delaware No Drawing. Continuation-impart of application Ser. No. 201,824, June 12, 1962, which is a continuation-in-part of application Ser. No. 805,020, Apr. 8, 1959. This application Feb. 7, 1964, Ser. No. 343,233 Claims priority, application Netherlands, Apr. 12, 1958, 226,774 6 Claims. (Cl. 260397.45)

This invention relates to a novel class of steroids wlu'ch exhibit important pharmacological properties and/or are useful as intermediates in the preparation of pharma cological active compounds.

This application is a continuation-in-part of our copending application Ser. No. 201,824, now Patent No. 3,198,792, which in turn is a continuation-in-part of Ser. No. 805,020 filed April 8, 1959 now abandoned.

All the known pharmacologically active steroids of the normal series which have a methyl substitute at the carbon atom 10 of the nucleus have this substituent in the 13 position. In general these compounds are characterized in having a substantially flat or planar molecular configuration of the nucleus, with the 1013 methyl group projecting from the plane of the nucleus.

It is now well accepted that the pharmacological ac-" tivity of the compounds of the normal series is attributable to the fact that the steroid molecule is capable of conforming to the configuration of the receptor. As a result, those skilled in the art believed heretofore that steroids of the normal series are highly stereo-specific in their interaction with the receptors. Since, in general, the nuclei of the pharmacologically active compounds of the normal series are substantially flat, it would be expected that the pharmacological activity is connected with a substantially flat structure of the nucleus and therefore it would not be expected that steroids having a substantially non-planar configuration of the nucleus would possess useful pharmacological properties.

A principal object of our invention is to prepare a new and novel class of steroids. This and other objects of our invention will be apparent from the description that follows.

Quite unexpectedly, we have discovered a novel class of steroids which behaves contrary to the aforesaid expectations in the steroid art. In contrast to the normal steroids wherein the methyl at the carbon atom 10 is in the position and the substituent at the carbon atom 9 is in the a position, in our novel class of steroids, the methyl substituent is in the a position while the substituent at the carbon atom 9 is in the ,8 position.

In addition to what has been mentioned above, with respect to the configuration of the 10 methyl group, and the substituent at the carbon atom 9, there is evidence to indicate that the structure of the nucleus of our novel compounds is non-planar. It is suggested that the nucleus of our novel steroids lies in two planes which intersect at a line drawn through the 8th and 9th carbon atoms of the nucleus. While this is a theory and we do not intend to be bound thereby, nevertheless, the indications are strong that the structure of the nucleus is non-planar. In view of the stereo specificity of the receptors, one would expect that such a deviation in the stereochemical structure of the steroid nucleus would render such steroids incapable of associating with the receptors. Consequently, it is surprising that our novel steroids possess any of the phar- 7 macological properties of the steroids of the normal series.

The steroids of our invention even posses a specificity which manifests itself in a fewer number of pharmacological activities than those possessed by the known corresponding compounds of the normal series, and/or exhibit new properties which are not possessed by the corresponding steroid of the normal series. In some cases, our novel steroids are unexpectedly superior in activities which are possessed in common with the corresponding compound of the normal series. Coupling the novel stereo-chemical configuration of the steroid nucleus with unexpected properties of our novel steroids, it is clear that an entirely new class of steroids has been discovered.

It is believed from all the experimental work accomplished to date that the specific 10a methyl 9/3 configuration of our novel steroids is the common dominant characteristic which produces the tendency for the highly specific actions thereof. Further, in this respect, the methyl, 9;? configuration of the nucleus in cooperation with the other substituents creates the particular properties possessed by our novel steroids. Hence, the configuration of the nucleus in our novel steroids is dominant with respect to the manner in which they differentiate from the steroids of the normal series.

Those novel steroids of the present invention in which the substituents at the carbon atoms 8, 9, 10, 13 and '14 have the same stereo-configuration as those in dihydroisolumisterone -(lumista-4,22-dien-3-one) are designed hereinafter as retrosteroids. Castells et al., Proc. of the Chemical Society, I an. 1958, page 7 has shown that dihydroisolumisterone has the configuration 8p, 9,8, 100: methyl, 13,6,

All the novel retrosteroids of the present invention exhibit highly specific pharmacological properties with respect to the following activities: parenteral progestational, oral progestational, pregnancy maintaining, deciduoma-inducing, fertility stimulating, anti-tumor, antiarteriosclerosis, fertility inhibiting, ovulation stimulating, ovulation inhibiting, anti-estrogenic, uterotrophic, antiuterophic, anabolic, renotrophic, anti-androgenic, pituitary stimulating, pituitary inhibiting, glucocorticoidal, anti-inflammatory, diuretic, etc.

With respect to the highly specific pharmacological propertes exhibited by the retrosteroids in comparison with the corresponding compounds of the normal series, it will be found that the retrosteroids have fewer or none of the properties in common with the corresponding compounds of the normal series and/ or the retrosteroids may have one or more pharmacological properties which are not possessed by the corresponding compounds of the normal series. In addition the retrosteroids can be unexpectedly superior in one or more of the pharmacological properties which are common to the corresponding compounds of the normal series. One of the outstanding advantages of our novel retrosteroids is that they can be used to produce a desired pharmacological activity with a substantially reduced number of side effects in comparison with the steroids of the normal series.

To illustrate the differences between our novel retrosteroids and those of the normal series, reference will now be had to comparisons for that purpose.

As previously stated, the retrosteroids of our invention are so different in the scope and specificity of their pharmacological properties that they are not comparable with their corresponding steroids of the normal series. The following examples will make these ditferences readily apparent.

The compound Epi of French Patent 1,091,734 has no glycogen storage activity and no anti-inflammatory activity while the corresponding retrosteroid of the invention has glycogen storage activity and is not anti-inflammatory.-

The 17a-methylandrosta-1,4-diene-17-ol-3-one of Brit- 3 ish Patent 750,834 is anabolic, non-parenteral progestational, and not pituitary inhibiting. The corresponding retrosteroid is anabolic, parenterally progestationally active and pituitary inhibting. I

Androst-4-ene-3,17-/8-diol of United States Patent 2,911,403 is androgenic, anabolic, not pituitary inhibiting, non-thymolytic, non-glucocorticoidal and is not antiinflammatory, whereas the corresponding retrosteroid is non-androgenic, non-anabolic, renotrophic, pituitary inhibiting, thymolytic, non-gluccorticoidal and is not antiinflammatory.

The 7-dehydroprogesterone of United States Patent 2,876,237 is progestationally active and metabolizes androgenic, while the corresponding retrosteroid is antiestrogenic, non-progestationally active, renotrophic and non-androgenic.

l7a-ethynylandrosta 4,6 dien-17-ol-3-one of United States Patent 2,882,282 is parenterally progestationally active, not orally progestationally active, anabolic, androgenic and renotrophic, whereas the corresponding retrosteroid is not parenterally progestationally active, orally progestationally active, non-anabolic, non-renotrophic, non-androgenic and anti-estrogenic. 6-dehydro-progesterone of United States Patent 2,882,282 has a decreased parenteral progestational activity in comparison to progesterone, is non-orally progestationally active and is metabolized androgenic, whereas the corresponding retrosteroid has an increased parenteral progestational activity in relation to retro-progesterone which by itself is more progestationally active than progesterone, is orally progestationally active and is non-androgenic even if incorrectly metabolized, 21 acetoxypregna 4,6 diene 3,20- dione of the same patent is weakly parenteral progestationally active, non-orally progestationally active, produces an increase in the Na/K ratio, has no glycogen storage activity, is not anti-inflammatory and is androgenic, whereas the corresponding retosteroid is orally and parenterally progestationally active, diuretic with no increase in Na/K ratio, non-androgenic, and is antiinflammatory. The pregna-1,4,6-triene-3,20-dione has no glycogen storage activity but is parenterally progestationally active but not orally progestationally active and is metabolized androgenic whereas the corresponding retrosteroid is not progestationally active but induces corpora lutea, is pituitary inhibiting and is non-androgenic even if metabolized incorrectly,

The 3,17/8-diacetoxyandrosta-3,S-diene of United States Patent 2,885,397 is androgenic, anabolic and is not pituitary inhibiting, whereas the corresponiing retrosteroid is non-androgenic, non-anabolic, renotrophic and pituitary inhibiting.

The activities which are mentioned above in connection with the compounds of the normal series are reported in the patents in which such compounds are disclosed.

Many additional comparisons based on our own experiments will be found in Table I following the examples.

In particular the novel steroids of our invention are the loot methyl, 9,8 steroids which contain at least 18 carbon atoms in the molecule and wherein any acyclic carbon chain when present and directly attached to the C carbon atom, in the 3 position, contains at least one and not more than five successive carbon atoms.

An important class of these novel steroids of our invention are those in which there is a substituent having a carbon atom directly attached to the C carbon atom which substituent is preferably in the ,8 position. In addition it is preferred that the substituent at the carbon atom 14 be in the a position and that the substituent at the carbon atom 8 be in the ,8 position.

indicated as being joined to the steroid nucleus by a broken line and a solid line placed side to side FORMULA I wherein R,, is a member of the group consisting of carbon to carbon double bonds present at the positions 1, 2, 3 and 4 and n,, is a whole number from 0 to 2;

R is a member of the group consisting of carbon to carbon double bonds present at the positions 5 and 6 and u is a whole number from 0 to 1;

R is a carbon to carbon double bond present at the position 11 and n is a whole number from O to 1;

R is a member of the group consisting of carbon to carbon double bonds present at one of the positions 15, 16 and 17 (20), and n is a whole number from 0 to 2;

R is a member of the group consisting of hydrogen, alkyl containing from 1-3 carbon atoms, hydroxy, acyloxy, aralkoxy, alkoxy and acylthio, and

n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, methylene, hydroxymethylene, alkoxymethylene, the methylene group --CH joining R and R hydroxy, acyloxy, alkoxy, aralkoxy, oxo, F, Cl and Br and n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, alkyl containing from 1 to 2 carbon atoms, substituted with hydroxy, 0x0 or both of the same, alkenyl containing from 2 to 5 3 carbon atoms, alkynyl containing from 2 to 3 carbon atoms, hydroxy, acyloxy, alkoxy and aralkoxy groups, the thio analogues of said hydroxy, acyloxy, alkoxy and aralkoxy groups, oxo, ketalized X0, F, Cl, Br, --NH alkyl alkyl -N and N H alkyl wherein each alkyl group contains from 1 to 3 carbon atoms and (2,3-d)-isoxazole, (3,2-c)-pyrazole, 2'-methyl (3,2 d)-thiazole and 2'-amino-(3,2-d)pyrimidine heterocyclic groups wherein each of said heterocyclics is formed by R together with carbon atoms 2 and 3 of the steroid nucleus and R and n is a Whole number from 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, hydroxy, acyloxy, aralkoxy, alkoxy, 0x0, F, Cl and Br and n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing fiom 1 to 3 carbon atoms, hydroxy, acyloxy, alkoxy, acylthio, aralkoxy,F, C1 and Br and n is a whole number from O to 1;

R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, methylene trifiuoromethyl, trichloromethyl, tribromomethyl, alkenyl containing from 2 to 3 carbon atoms, alkynyl containing from 2 to 3 carbon atoms, hydroxy, acyloxy, alkoxy, aralkoxy, acylthio, oxo, nitro, amino, F, Cl and Br and n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing 1 to 3 carbon atoms, hydroxy, acyloxy, acylthio, alkoxy, aralkoxy, oxo, amino, F, C1 and Br, and

n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, hy-

droxy, acyloxy, alkoxy and aralkoxy and R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, hydroxy, acyloxy, alkoxy, aralkoxy, F, Cl and Br and R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, methylene, alkenyl containing from 2 to 3 carbon atoms, alkynyl containing from 2 to 3 carbon atoms, hydroxy, acyloxy, alkoxy, aralkoxy, 0x0, amino, F, Cl and Br and n is a whole number fiom 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3 carbon atoms, hydroxy, acyloxy, alkoxy, aralkoxy, oxo, F, Cl and Br and n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, methla a yl, hydroxymethyl, formyl and together with R the radical 11 is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, alkyl containing from 1 to 3-carbon atoms, methylene, trifluoromethyl, trichloromethyl, tribromomethyl, alkenyl containing from 2 to 3 carbon atoms, alkynyl containing from 2 to 3 carbon atoms, hydroxy-methylene, hydroxy, acyloxy, alkoxy, aralkoxy, oxo, amino, F, Cl and Br and n is a whole number from 1 to 2;

R is a member of the group consisting of hydrogen, hy-

droxy, acyloxy, alkoxy, aralkoxy, thio-analogues of said hydroxy, alkoxy and aralkoxy groups, alkyl containing from 1 to 6 carbon atoms, alkenyl containing from 2 to 6 carbon atoms, alkynyl containing from 2 to 6 carbon atoms, and said alkyl, alkenyl and alkynyl groups substituted with at least one member of the group consisting of hydroxy-, hydroxy esterified with inorganic acid, hydroxy esterified with inorganic acid of which one of the hydrogen atoms is replaced by an alkali metal atom, acyloxy-, alkoxy-, aralkoXy-, oxo-, amino-, F-, Cl-, Brand the thio analogues of said hydroxy, alkoxy and aralkoxy groups and R' is a member of the group consisting of hydrogen, hydroxy, acyloxy, alkoxy, aralkoxy, thio-analogues of said hydroxy, alkoXy and aralkoxy groups, F, Cl, Br, alkyl containing from 1-6 carbon atoms, alkenyl containing from 26 carbon atoms, alkynyl containing from 26 carbon atoms, said alkyl, alkenyl, and alkynyl groups substituted with at least one member selected from the group consisting of hydroXy-, acyloXy-, alkoXy-, ara1koxy-, the thio-analogues of said hydroxy, alkoxy and aralkoxy groups, carboxy-, oXo-, amino-, F, Cl and Br, the oxo group joining R and R' the group CH joining R and R and a spirolactone containing from 3-6 carbon atoms joining R and R' a ketal group joining R and R and a ketal group joining R' q and R An interesting group of the hormonal retrosteroids of our invention are the 10:! methyl steroids corresponding to the general structural formula:

FORMULA II acne CH3 17) llh'l R is a substitutent selected from the group consisting of n is a whole number from 1 to 2;

R is a substituent selected from the group conslstlng of keto, hydroxy, esterified hydroxy and etherified hydroxy radicals and n is a whole number from .1 to 2;

R is a substituent selected from the group consisting of hydrogen, chlorine and fluorine and n is a whole number from 1 to 2;

R is a substituent selected from the group consisting of hydrogen, bromine, chlorine, fluorine, hydroxy, methyl, etherified hydroxy and esterified hydroxy radicals and n is a whole number from 1 to 2;

R is a substituent selected from the group consisting of hydrogen, bromine and fluorine and R is a substituent selected from the group consisting of hydrogen, the hydroxy radical and keto radicals and fl is a whole number from 1 to 2;

R is a substituent selected from the group consisting of hydrogen, hydroxy, etherified hydroxy, esterified hydroxy and ethyl and methyl radicals and n is a whole number from 1 to 12;

R and R' are each selected from the group consisting of hydrogen, aliphatic hydrocarbon radicals containing from 1-6 carbon atoms, hydroxy derivatives of said aliphatic hydrocarbon radicals, etherified hydroxy derivatives of said aliphatic hydrocarbon radicals, esterified hydroxy derivatives of said aliphatic hydrocarbon radicals and keto derivatives of said aliphatic radicals, and jointly the keto radical with the proviso that at least one of the substituents defined by R and R q being a substituent other than hydrogen.

R is a carbon to carbon double bond present at at least one of the positions 1, 2, 3, 4 and n, is a whole number from -2;

R is a member of the group consisting of carbon to carbon double bonds present at the positions 5, 6, 7 and 8 and In, is a whole number from 0 to 2;

R is a member of the group consisting of carbon to carbon double bonds present at the positions 8 (14) and 9 (11) and n is a whole number from 0 to 2;

R is a member of the group consisting of carbon to carbon double bonds present at one of the positions 15 and 16 and n is a whole number from 0 to 2.

A particular group of compounds of Formula II are those in which there is a conjugated carbon double bond system at the carbon atoms and 7 of the steroid nucleus.

Referring to the general Formula I, three particular groups of retrosteroids having female hormonal activity are those in which all substituents are the same as defined therein except that:

Group A R17 is Group B R is OR wherein R is hydrogen, alkyl or acyl and R is alkyl of 2-6 carbon atoms, alkenyl or alkynyl each of 26 carbon atoms.

Group C R17 is (|f C H;

and

8 trophic, anti-uterotrophic, anti-estrogenic, fertility stimulating, fertility inhibiting, anti-arteriosclerosis, corpora lutea induction, ovulation inhibiting and/ or ovulation stimulating activities. In addition some of these compounds may have anti-tumor, pituitary stimulating and/ or pituitary inhibiting activities.

As to the compounds falling within the groups A, B and C, the introduction of a double bond at the carbon atom number 1 has a tendency to decrease progestational activity and to enhance ovulation stimulating activity. The presence of a keto group at carbon atom number 3 and a double bond at carbon atom 4 has a tendency to enhance oral and parenteral progestational activity as compared with the same activity of the corresponding compound of the invention in which such a keto atom and such a double bond are absent.

The presence of a keto group at carbon atoms number 3 and double bonds at carbon atoms numbers 4 and 6 has a tendency to enhance oral and parenteral progestational activity and anti-uterotrophic activity as compared with the same activities of the corresponding compound of the invention in which such a keto group and such double bonds are absent. The presence of an alkoxy or acyloxy group at carbon atom number 3 and double bonds at carbon atoms 3 and 5 have a tendency to decrease the intensity of progrestational activity but prolong the same as compared with the same activities of the corresponding compound of the invention in which such alkoxy, acyloxy, and double bonds are absent. A fluoro or chloro atom at carbon atom number 6 has a tendency to increase any female hormonal activity already possessed by the compound. A double bond at carbon atom number 7 has a tendency to decrease any female hormonal activity already possessed by the compound but to enhance antiarteriosclerotic activity.

Referring to the Formula I three particular groups of retrosteroids having male hormonal activity are those in which all substituents are as defined therein except that Group D R is hydrogen, or OR, wherein R is hydrogen or acyl, and R37 is hydrogen or 0R, wherein R is hydrogen or acyl, provided that R and R' are neither both hydrogen nor both 0R.

Group E R and R' together form the 0x0 Group F R is OR wherein R is hydrogen, alkyl or acyl, and RIM is methyl.

The compounds falling within the above groups D, E and F possess at least one of the following pharmacological activities, namely, anti-androgenic, anabolic, renotrophic, anti-catabolic, catabolic, increase or decrease of spermatogenesis and/0r anti-anemic activities.

In addition some of these compounds may have antitumor, pituitary stimulating and/or pituitary inhibiting activities.

With regard to the compounds falling within the groups D, E and F the combination of a keto group at the carbon atom number 3 with a double bond at carbon atom number 4, has a tendency to enhance anabolic and/0r renotropic activities. The combination of a keto group at carbon atom number 3 with double bonds at carbon atoms 4 and 6 in the compounds falling within the groups D, E and F tends to impart anti-androgenic and/or pituitary inhibiting activities. A halogen atom especially a fluorine or chlorine atom at the carbon atom number 6 in compounds falling within the group D-F has a tendency to increase the anabolic activity.

Referring to Formula I, particular groups of retro- 9 steroids having cortiocoidal activity are those in which all substituents are as defined therein except that Group G R17 is o CHaOR and R is hydrogen;

Group H R17 is o -CH2OR and R is OR;

Group 1 One of the substituents at carbon atom 11 is hydroxy. R17 is o BOHzOR and R lq is OR;

Group J The substituent at carbon atom 11 is 0X0, R is El-CHzOR and R'm is OR and wherein in these groups G, H, I, J, R is hydrogen or acyl The compounds falling with the G, H, I and J groups possess at least one of the following properties, namely, gluco-corticoid, anti-gluco-corticoid, anti-inflammatory, mineralo-corticoid, anti-mineralo-corticoid, diuretic, antidiuretic, anti-allergic, and/ or anti-diabetic activities.

In addition some of these compounds may have antitumor, pituitary stimulating and/or pituitary inhibiting activities.

The presence of a keto group at carbon atom number 3 and an olefinic double bond at carbon atom number- 4 has a tendency to enhance one or more of the corticoidal activities of the retrosteroids falling within the groups G, H, I and I as compared with the corresponding activities of the corresponding compounds of the invention in which such a keto group and such a double bond is absent. Introduction of another olefinic double bond at carbon atom number 6, in addition to the keto group at carbon atom number 3 and the double bond at carbon atom number 4, has a tendency to enhance the corticoidal activity still further as compared with the corresponding activities of the corresponding compound of the invention in which such a double bond is absent.

In general, the activity of the retrosteroids of the groups A-J inclusive of the present invention show a tendency to be enhanced by the introduction of a double bond at carbon atom number 6, as compared with the corresponding activity of the corresponding compound of the invention in which such a double bond is absent.

Among the representative retro-steroids of our invention are the following:

218,21-difiuoro-retro-progesterone 1-dehydro-2,21-difluoro-retro-progesterone 1 l-oxo-retro-progesterone 1 1fi-hydroXy-retro-progesterone 1,6-bisdehydro-6-methoxyretro-progesterone 6,8-methoxy-retro-progesterone 1-dehydro-6/3-methoxy-retro-progesterone 1-dehydro-6fi-fluoro-21-acyloxy-retro-progesterone 6-dehydro-6-methoxy-21-acyloxy-retro-progesterone 6B-methoXy-21-acyloxy-retro-progesterone l-dehydro-Zl-acyloxy-retro-progesterone 6fi-fluoro-21-acyloXy-retro-progesterone 16a-hydroxy-21-acyloxy-retro-progesterone 3-enolether of 21-acyloxy-retro-progesterone 3-enolacylate of 21-acyloxy-retro-progesterone 3-desoxy-6-dehydro-retro-progesterone retro-pregna-4,6-dien-20-ono-(2,3-d)-isoxazole retro-pregna-4,6-dien-20-ono- 3,2-c -pyrazole 6-dehydro-6-fiuoro-retro-progesterone 1,6-bisdehydro-6-fiuoro-retro-progesterone 1,fi-bisdehydro-6-chloro-retro-progesterone 6-dehydro-2 1-fluoro-retro-progesterone 1,6-bisdehydro-2l-fluoro-retro-progesterone 6-dehydro-21,21-difluoro-retro-progesterone 1,6-bisdehydro-2,2l-difluoro-retro-progesterone 6-dihydro-16a, 17 a-dihydroxy-retro-progesterone '16, 17-

ketal 6-chloro-6-dehydro-16a,17a-dihydroxy-retro-progesterone 16,17-ketal 6-chloro-6-dehydro-16a,17a-dihydroxy-retro-progesterone 6-dehydro-16a, 17a-dihydroxy-retroprogesterone 6-dehydro-6-fluoro-21-acyloxy-retro-progesterone 1,6-bisdehydro-21-acyloxy-retro-progesterone 6-dehydro-16a-hydroxy-21-acyloxy-retro-progesterone 23,21-difluoro-6-dehydro-retroprogesterone 6a-chloro-retro-testosterone 17-acylate 1-dehydro-2-methyl-retro-testosterone 13 1-dehydro-2-methyl-retro-testosterone 17-acylate (13) 4-methyl-retro-testosterone 3-enolacylate of 4methyl-retro-testosterone 17-acylate 4-methyl-6-del1ydro-retro-testosterone 4-methyl-6-dehydro-retro-testosterone 17-acylate 4-chloro-retro-testosterone 4-chloro-retro-testosterone 17-acylate 4-bromo-retro-testosterone 4-hydroxy-retro-testosterone 4-hydroXy-6-dehydro-retro-testosterone 2 3-fluoro-retro-testosterone (16) 17a,21-diacyloxy-retro-progesterone 3-enolacylate of 17a,2l-diacyloxy-retroprogesterone Git-fluoro-17a,21-diacyloxy-rtroprogesterone 6-dehydro-6-rnethoxy-170:,2l-diacyloxy-retro-progesterone 1,6-bisdehydro-17a-acyloxy-retro-progesterone 1,6-bisdehydro-1-methyl-17a-acyloxy-retro-progesterone 3-desoxy-6-dehydro-17a-acy1oXy-retro-progesterone 3-enolether of 6-dehydro-17a-acyloxy-retro-progesterone 6-chloro-6-dehydro-17a-acyloxy-retro-progesterone 6-dehydro-6-fluoro-17a-acyloxy-retro-progesterone LG'blSdGhYdIO-G-fiUOI'O-I70t-3CY10XY-ICtl'0PIOgSt6I'OI16 1,6-bisdehydro-6-chloro-17a-acyloxy-retro-progesterone 6-dehydro-17a,21-diacyloXy-retro-progesterone 6-dehydro-6-fluoro-17a,21-diacyloxy-retro-progesterone 6-chloro-6-dehydro-1711,21-diacyloxy-retrmprogesterone 16m-methyl-retro*progesterone 16,3-methyl-retro-progesterone 6-dehydro-1 6a-metl1yl-retro-pro gesterone 6-dehydro-16a-methyl-17u-acyloxy-retro-progesterone 16-methylene-17a-acyloxy-retro-progesterone 3-enolacylate of 1Got-methyl-l7a-acyloxy-retro-progesterone 16a-methyl-17a-acyloXy-retro-progesterone 16;3-methy1-17a-acyloXy-retro-progesterone 6-dehydro-16-methylene-17a-acyloxy-retro-progesterone 16a, 17a-dihydroxy-retro-progesterone 1 6, l7-ketal 16a-methyl-21-acyloxy-retro-progesterone 16a,17a-dihydroXy-21-acyloxy-retro-progesterone 16,17-

ketal 6,8,16a-dimethyl-17u-acyloxy-retro-progesterone 6 -dehydro-6, 1 6a-dimethyl- 17u-acy1oxy-retro -progesterone 6-dehydro-6, 1 6a-dimethyl-retro-progesterone 16u-methyl-retro-progesterone S-enolacylate of l7a-methyl-retro-progesterone 6-dehydro-17a-methyl-retro progesterone 17rx-brorno-retro-progesterone 1-dehydro-17u-bromo-retro-progesterone 15 uents except that rz is a whole number from to 2, R' is hydrogen and R is the group QV wherein Q is an alkyl of 1-3 carbon atoms, phenyl, the group wherein R and R are both alkyl or the cyclic group wherein R" and R' form together a bivalent hydrocarbon radical and Q is phenyl, hydrogen or alkyl of 1-3 carbon atoms with the proviso that Q is hydrogen when Q contains nitrogen and Q is an alkyl group of 1-3 carbon atoms but not more than three carbon atoms together with Q when Q is alkyl.

Another group of these novel intermediates in those which also have the same structure as Formula I in respect to all the substituents except that n is a whole number from 0 to 2 and R together with R is the group wherein X is the carbon atom 17 of the nucleus.

Other novel intermediates are those similar to the above two groups and Formula I but in which there are present a double bond at 8(9) or 9(11) and/ or one or more epoxy groups joining the following pairs of nuclear carbon ,atoms: 1, 2; 4, 5; 5, 6; 6, 7; 9, ll; 11, 12; 14, 15 and 16, 17.

The use of these novel steroids of our invention in the preparation of our novel hormonally active retro-steroids will be apparent from the description of the methods of preparing these retro-steroids which follows:

We may start with steroids already containing the 100: methyl 9,9 configuration such as lumisterol 2 or lumisterol 3.

Also the u-methyl, 9,8 configuration of the retro-steroids of our invention may be prepared by irradiation with ultraviolet light of certain normal steroids having a A configuration in the same manner that ergosterol and 7-dehydrocholesterol are converted into lumisterol 2 and lumisterol 3, respectively.

Thus, irradiation (preferably with ultraviolet light) of 5,7-bisdehydro-normal steroids, having the IO-methyl group in 5 position, a hydrogen atom at carbon atom 9 in a position, a substituent at carbon atom 13 in [3 position and a substituent at carbon atom 14 in on position, followed by eliminations of the double bond between carbon atoms 7 and 8 produces steroids having a nucleus of which the configuration at the carbon atoms 10, 9, 8, 13 and 14 is that of the nucleus of our novel retro-steroids (10a methyl, 9;), 135, 14a).

In this manner, the loot methyl, 9,8 configuration in the novel steroids of our invention, can be introduced as set out in the foregoing paragraph. The starting materials used in the irradiation process may already contain the substituents as set out in Formula I. However, sometimes the presence of certain light absorbing chemical systems other than the 5 ,7-bisdehydro system may disturb the conversion into our novel 10oz methyl, 9;? steroids.

In principle this method of introducing the 1001 methyl, 95 configuration by irradiation with ultraviolet light may be applied to all normal steroids (those containing a 10,8 methyl, 9a configuration) with two limitations. One of these limitations is that there be no substituent present providing steric hindrance of the lO-methyl group. It is particularly important that the substituents present on the 1, 9 or 11 positions of the steroid molecule do not stericly hinder the IO-methyl group. Also there must be no substituents which have a natural absorption between 220 and 330 m, or materially influence the absorption of the A system. Thus no non-ketalized keto group or one or more double bonds in conjugation with the A system may be present.

Starting products for the preparation of the retrosteroids according to the invention may be, for example, those natural sterols which have a saturated or unsaturated aliphatic car-bon chain, such for example ergosterol.

Alternatively, the starting products may be found in the classes of the sapogenines, which have a heterocyclic group in the side chain, for example, diosgenine. In both cases the M -system should be provided for the radiation if it is not already present in the molecule.

The introduction of the 10m methyl, 9;? configuration may be the first step to be carried out as in the following general method:

(a) Introduction of the retro-structure into steroids of the normal series by irradiation of suitable A steroids; (b) Introduction of substituents into retro-steroids;

(c) A series of reactions to obtain a desired grouping at carbon atom 17, starting from retro-steroids with an aliphatic side chain of 6-10 carbon atoms at carbon atom 17.

The sequence of the processes as described may be varied. For example, starting irom a retro-steroid which has been prepared beforehand, for example dihydroisolumisterone, the side chain, degradation and introduction of the desired substituents may be carried out entirely or partially in the inverted sequence, 2-fluororetro-progesterone may be prepared for example by introducing a fluorine atom in the 2-position in retroprogesterone or by side chain degradation of 2-fiuorodihydroisolumisterone.

'It is also possible to carry out the irradiation with steroids of the normal series, in which all of the desired substituents have been introduced, or after entire or partial side chain degradation for example with 16- methyl-pregna-5,7-diene-3[i-ol-20-one.

The sequence of the processes which is chosen can be decided separately for the product and starting materials employed.

Changes in the nuclear structure other than the introduction of the s methyl, 913 configuration may be carried out as follows? 14B-steroids may be prepared by hydrogenation of 14(15) unsaturated steroids, for instance with hydrogen and using palladium as a catalyst. Thus pregna-5,l4,16- trien-3 fi-ol-20-one when hydrogenated yields 145,175- pregna-5-en-3fi-ol-20-one. (Fieser and Fieser, Steroids 1959, page 567.)

t-SI61'OldS may be prepared by irradiation of 17-keto steroids with ultra violet light according to Fieser and Fieser Steroids, 1959, page 520. Thus irradiation of 3a-hydroxy 5a androstan-17-one yields 3a-hydroxy-5a, 13a-androstan-17-one.

80c steroids may be prepared by hydrogenation of 11-- keto-A steroids in the presence of a catalyst. (Tetrahedron 1, 22 (1957).) Thus 3B-hydroxy-1 l-oxoand rost- B(9)-ene may be hydrogenated in the presence of palladium to produce 313-hydroxy-11-oxo-8a-.androstane.

18-nor steroids may be prepared by reacting 18-hydroxy-17-keto steroids with sodium hydroxide (Fieser and Fieser Steroids 1959, page 470). Thus by reacting 3B, 18-dihydroxy-androst-5-en-17-one with sodium hydroxide will be produced 3B-hYdIOXY-1S-IIOFEIIldIOSlI-S-(ill-17-0116. The expressions: inorganic acid, acyl, acyloxy, alkoxy,

aralkoxy, acylthio, ketol and ketalized oxo, whenever used in this application, have the following meanings.

Inorganic acid is one of the following inorganic acids: phosphoric acid, sulphuric acid, nitric acid, nitrous acid, boric acid.

Acyl or acyloxy are the acyl or the acyloxy groups respectively of the following acids:

(a) Saturated or unsaturated, branched or unbranched, cyclic or non-cyclic aliphatic monoor dicarboxylic acids having 120 carbon atoms wherein the aliphatic part of the molecule may be substituted by one or more halogen atoms, amino groups, sulphonic acid groups;

(b) Phenylalkyl monoor dicarboxyl-ic acids of which the phenyl part of the molecule may be substituted with alkyl groups containing 1-3 carbon atoms, sulphonic acid groups, alkoxy groups of which the alkyl group is branched or unbranched and contains 1-10 carbon atoms and acids of which the alkyl part of the phenyl alkyl monoor dicarboxylic acid contains from -6 carbon atoms and is branched or unbranched, saturated or unsaturated. Of these acids the following specific examples may be given: formic acid, acetic acid, acroleic acid, isobutyric acid, palmitic acid, cyclohexane-mono-carboxylic acid, trichloroacetic acid, aminoacetic acid, oxalic acid, malonic acid, maleic acid, benzoic acid, terephthalic acid, p-ethyl benzoic acid, benzene sulphonic acid, m-ethoxy benzoic acid, phenyl acetic acid, and cinnamic acid.

Alkoxy is the alk-oxy group of mono or di-aliphatic alcohol of which the alkyl group contains from 1-20 carbon atoms and which may be branched or unbranched, cyclic or non cyclic, saturated or non saturated and which alkyl group may be substituted with halogen or alkoxy, whereby this latter alkoxy substituent may be attached to the same carbon atom of the steroid nucleus as the main alkoxy group. Of these alcohols the following examples may be given: methanol, ethanol, propanol-Z, Z-chloro ethanol-1, 2-ethoxy ethanol-1 and glycol.

Aralkoxy is the aralkoxy group of a mixed aromatic aliphatic alcohol in which the hydroxyl group is attached to the aliphatic part of the molecule and of which the alkyl (ene) group contains from 06 carbon atoms which may be branched or unbranched. Of these alcohols the following examples may be given: benzyl alcohol and phenol.

Acylthio is the acylthio group of acyl thiols. In acylthio the expression acyl has the meaning as given hereabove.

A ketal is the dihydroxy steroid ketal of an aliphatic aldehyde. aliphatic ketone, mixed aliphatic aromatic aldehyde, mixed aliphatic aromatic ketone, or a diaromatic ketone.

Ketalized oxo is the ketalized oxo group resulting from the reaction between two molecules of a monohydroxy aliphatic alcohol containing from 1 to 6 carbon atoms and one molecule of an oxo group containing a,9 8- methyl steroid, or resulting from the reaction between One molecule of a dihydroxy aliphatic alcohol containing from 1 to 6 carbon atoms and one molecule of an oxo group containing 10a-methyl-9B-steroid.

The introduction of alkyl substituents may be carried out as follows:

(1) By the reaction of keto-, conjugate ketoand doubly conjugate keto-steroids with alkylation agents, such as, for example, Grignard reagents, alkyl-alkali-metal compounds or an alkyl halide, in the presence of an alkali metal alkoxide.

For example, according to the method described by Zderic et al. (I. Am. Chem. Soc., 82, 3404 (1960)) 11- keto-retro steroids lithium give the corresponding 11- methyl11-hydroxy steroids. In this manner, ll-keto-retroprogesterone 3,20-bisethylene ketal may be converted to the corresponding ll-methyl-ll-hydroxy compound. After acid hydrolysis, this provides ll-methyl-ll-hydroxy-retroprogesterone.

Further, l1-keto-retro-testosterone 3-ethylene ketal may The reaction of 3-keto-retro steroids with methyliodide in the presence of potassium-t-butoxide by the procedure of Ringold et al. (J. Am. Chem. Soc., 81, 427 (1959)) provides the corresponding 2,2-dimethyl steroids. In this manner, 2,2-dimethyl-retro-pregnane-3,ZO-dione 20-ethylene ketal may be produced from retro-pregnane-3,20- dione 20-ethylene ketal.

Further, 2,2-dimethyl-retro-androstane 17,6-ol-3-one may be produced from retro-androstane-l7B-ol-3-one.

Addition of methyl Grignard reagents to A -20-ketoretro-pregnanes according to the method described by Bernstein ct al. (J. Org. Chem., 26, 269 (1961)) provides 16-methyl compounds. Thus, 3B-hydroxy-16-methyl-retropregn-S-ene-ZO-one may be produced from 3B-acetoxyretro-pregna-5,l6-diene20-one.

Conversion of 3-keto-A -retro-ster0ids with methyl halides in the presence of potassium-t-butoxide by the pro cedure of Woodward et al. (I. Am. Chem, Soc., 76, 2852 (1954)) gives 3-keto-4,4-dimethyl-M-steroids. For example, such a reaction with bismethylene-dioxy-retro-hydrocortisone gives the 4,4-dimethyl-l7(20),20-bismethylenedioxy-retro-pregn-S-ene-3,20-dione.

Further, with retro-testosterone, such a reaction gives the 4-4, dimethyl-retro-androst-S-ene-17B-ol-3-one.

(2) By the addition of diazomethane to double bonds, succeeded by pyrolysis or cleavage under acid conditions of the pyrazolines, if required with subsequent hydrogenation of the compounds obtained. According to the procedure of Wiechert and Kaspar (Chem. Ber., 93, 1710 (1960)) A -3-keto-retro-steroids react with diazomethane with the formation of pyrazolines, from which the 1,2-methylene-A -3-keto-steroids are produced by pyrolysis. Under the action of, for example, acid alumina, however, the pyrazolines are converted into the A I-methyl steroids. For example, 17a-acetoxy-1,Z-methylene-retropregna-4,6-diene,3,20-dione and 17a-acetoxy-1-methyl-retro-pregna-1,4,6-triene-3,20-dione may be produced from 17a-acetoxy-retro-pregna-1,4,6-triene-3,20-dione.

Further, 1,2-methylene-retro-androsta-4,6-diene--01- 3-one 17-acetate and 1-methylretro-androsta-1,4,6-triene- 17fl-ol-3-one 17-acetate may be produced from retroandrosta-l,4,6-triene-17B-ol-3-0ne 17-acetate.

(3) By the reaction of a methylene group activated by a keto-group, with a monoor dicarboxylic acid ester where R may be H or succeeded by a reaction with an alkyl halide with subsequent splitting off of the carboxylic acid group.

For example, 3-keto-A -retro-steroids when reacted with diethyloxalate in the presence of sodium hydride according to the method described by Ringold et a]. (J. Am. Chem. Soc., 81, 427 (1959) after reaction with a methyl 19 halide followed by a reaction with an alkali-alkoxide give the 2-methyl-3-keto-A -retro-steroids Thus, from retropregn-4-ene-20-ol-3-one 20 acetate the 2-methylretropregn-4-ene-20-ol-3-one ZO-acetate may be produced. I

Further, in this manner 2-methyl-retro-testosterone may be produced from retro-testosterone.

(4) By the reaction of retro-steroid epoxides with, for example, alkyl Grignard reagents. For example, 3-cycloethylenedioxy-5(6) epoxy retro steroids after reaction with methyl magnesium halides according to the method described by Babcock et al. (J. Am. Chem. Soc., 80, 2904 (1958)) give the 6-methyl-5-hydroxy compounds, from which after hydrolysis and dehydration the 6-methyl-3- keto-M-retro-steroids can be produced. Such a reaction with, for example, 5,6-epoxy-17a-hydroxy-retro-pregnane- 3,20-dione bisethylene acetal gives the corresponding 5- hydroxy-6-methyl compound which, after hydrolysis and subsequent dehydration, provides the 6-methyl-17a-hydroxy-retro-progesterone.

Further, from 5 ,6-epoxy-retro-androstane-3, l 7-diol may be obtained in this manner S-hydroxy-6-methyl-retro-androstane-3,17-diol.

The introduction of double bonds in retro-steroids may be effected by the following methods:

( 1) By microbiological dehydrogenation, as described,

for example, for the production of l-dehydro-steroids by Sih et al. (J. Am. Chem. Soc., 82, 2653 (1960)). In this manner, retro-progesterone may be converted into l-dehydro-retro-progesterone, or, retro-testosterone may be converted into 1-dehydro-retro-testosterone.

(2) By the direct dehydrogenation of saturated or nonstatur ated steroid ketones and of unsaturated steroids with, for example, mercury acetate, iodine pentoxide, selenium dioxide, manganese dioxide, substituted quinones such as chloranil and dichlorodicyanobenzoquinone, for example by the methods of Heilbron et al. (J Chem. Soc., 1935, 1221), Burn et al. (Proc. Chem. Soc. 1960, 14), Agnello et al. (J. Am. Chem. Soc., 82, 4293 (1960)) and Sondheimer et al. (J. Am. Chem. Soc., 75, 5932 (1 953)). In this manner, 6-dehydro-retro-progesterone may be produced from retro-progesterone and 1,6-bisdehydro-17a-acetoxy-retro-progresterone may be produced from 6-dehydro-17 t-acetoxy-retro-progresterone. 3,17-diacetoxy-retro-androsta-S,7,9(11)-triene may be obtained from 3,l7-diacetoxy-retro-androsta-S,7-diene, and 1-dehY- droretro-testosterone may be obtained from retro-testosterone.

(3) By the splitting off of substituents, for example dehydrohalogenation of halogenated retro-steroids, for

example with an organic base, as has been described by Djerassi' et al. (J. Am. Chem. Soc., 72, 4534 (1950)) for the production of 6-dehydrofrom 6-halo steroids. In

this manner, 6-dehydro-retro-progesterone may be produced from 6-bromo-retro-progesterone. Further, 6-dehydro-retro-testosterone 17-acetate may be produced from 6-bromo-retro-testosterone 17-acetate.

Another example of splitting off is the dehydration of hydroxylated retro-steroids, for example under the influence of phosphorus-oxychloride and an organic base, from methane sulphonyl chloride or from methyl chlorosulphite, for example by the methods described by Chamberlin et al. (J. Org. Chem., 25, 295 (1960)). In this manner, retro-hydrocortisone acetate may be converted into l7u,21-dihydroxy-retro-pregna-4,9( 11 )diene-3,20-dione 2l-acetate. Further, 11-hydroxy-retro-androst-4-3n3- 3,20-dione may be converted into retro-androsta-4,9(11)- diene-3,20-dione.

The introduction of hydroxy groups into retro-steroids may be eifected as follows:

(1) By microbiological hydroxylation, for example as described by Peterson et al. (J. Am. Chem. Soc., 74, 1871 (1952)). In this manner, 11,17m,21-trihydroxy-retropregn-4-ene-3,20-dione may be produced by microbiological hydroxylation of 1711,21-dihydroxy-retro-pregn-4-ene- 3,20-dione. Further, 11-hydroxy-retro-testosterone may 20 I be produced by microbiological hydroxylation of retrotestosterone according to the same method.

(2) By cleavage of epoxy-retro-steroids. This may be effected by means of reducing agents, such as, for example, alkali-bariumand alkali-aluminum-hydride, chromium salts, and by catalytic hydration. According to the method described by Cole et al., (J. Org. Chem., 19, 131 (1954)), the 16 hydroxy compounds may be produced from 16,17-epoxy-retro-steroids. Thus, 16,17-epoxy-retropregn-4-ene-2-1-ol-3,20-dione yields the retro-pregn-4-ene- 16,21-diol-3,20-dione. Further 3,17-dihydroxy-5(6)-e'poxy-retro-androstane may be converted into 3,6,17-trihydroxy-retro-androstane.

Cleavage of epoxy-retro-steroids may also be performed by means of hydro halogenic acids, for example as described by Ringold et al. (J. Am. Chem. Soc., 78, 816 (1956)) for the production of 17u-hydroxy-pregnames from 16,17-epoxy-pregnanes. According to this method, 16,17-epoxy tretro+progesterone, after reaction with hydrogen bromide succeeded by reductive debromination, yields the 17a-hydroxy-retro-progesterone. Further, 9 1 1 )-epoxy-retro-androst-4-ene-17fl-ol-3-one with hydrogen fluoride yields 11-hydroxy-9-fluoro-retro-testosterone.

(3) By hydroxylation of double bonds, for example with osmium tetroxide. For example, osmate esters, produced by oxidation of alkenes with osmium tetroxide, are split up into the diols, employing the procedure of Baran (J. Org. Chem. 25, 257 (1960)). In this manner, 3-hydroxy-retro-pregn-S-ene-ZO-one may be converted into 3,5,6-trihydroxy-retro-pregnane-ZO-one. Further, 3,8-hydroxy-retro-androst-S-ene-l7-one may be converted into 3B,5,6-trihydroxy-retro-androstane-17-one.

(4) By the reaction of enol ethers or enol esters with organic peracids. If, for example, A -3-enol acylates are reacted with per-acids, employing the methods of Rome et al. (J. Org. Chem, 19, 1509 (1954)), the 3-keto- A -6-hydroxy steroids are produced. T'hus, 3-acetoxyretro-pregna-3,5-diene-20-one with monoperphtalic acid yields the 6-hydroxyaretro-progesterone. Further, 3,175- diacetoxy-retro-androsta-3,S-diene with monoperphtalic acid yields the 6-dehydro-retro-testosterone 17-acetate.

(5) By the hydrolysis or acyloysis of halo-steroids, for example with an alkali salt of a carboxylic acid by the method of Ruschig (Ber., 88, 878 (1955)), 21-iodosteroids being converted with potassium acetate into the 21-acetoxy compounds. Hydrolysis of these compounds, for example according to Robinson (J Am. Chem. Soc. 82, 4611 (1960)) with perchloric acid yields the free 21-hydroxy compounds. The hydrolyses may suitably be performed with a solution of potassium bicarbonate in aqueous methanol. Thus, 21-iodo-retro-progresterone may be converted into 21-acetoxy-retro-progesterone, which again may be converted by hydrolysis into 21-hydroxyretro-progesterone. Further, in this reaction 6-bromoretro-testosterone 17-acetate gives Z-acetoxy-retro-testosterone 17-acetate, from which, after hydrolysis, 2-hydroxy-retro-testosterone is obtained.

(6) By reduction of keto-steroids, for example with lithium aluminum hydride by the method described by Sondheimer et al. (J. Am. Chem. Soc., 75, 5930, 5932 (1953)). In this manner, retro-progesterone may be reduced to retro-pregn-4-ene-diol. Further, for example, retro-androst-4-ene-3,17-dione may be reduced to retroandrost-4-ene-3,17-diol.

(7) By hydrolysis of esters or ethers, for example by the method of Dory et al. (C.A., 53, 17181 (1955)) with sodium methylate in methanol. The saponification may also be,performeds ,w ith dilute inorganic acids, dilute inorganic, bases, sodium carbonate and sodium bicarbonate. Thus, hydrolysis of 1704,21-dihydroxy-retro-progesterone 21-acetate with sodium bicarbonate in dilute dioxane gives the 17a,21-dihydroxy-retro-progresterone. Further, the hydrolysis of retro-test0sterone l'Z-acetate gives retrotestosterone.

The introduction of acyloxy groups into the retrosteroid series may be carried out as follows:

(1) By acyloysis of halo-retro-steroids as described hereinbefore.

(2) By the reaction of hydroxyand keto-retro-steroids with acids, acid anhydrides or acid chlorides in the presence of, for example, a catalyst (for example, ptoluene sulphonic acid, pyridine-HCl) or acid binding reagents (for example organic bases) or water-binding reagents (such as, for example, trifiuoro acetic acid anhydride). For example, 17methyl-retro-testosterone may be esterified with the acid chloride of B-phenyl-propionate in pyridine by the method of Gould et al. (J. Am. Chem. Soc., 79, 4472 (1957)) to form 17-(B-phenyl-propionate) ester of 17a-ethylretro-testosterone. Further, retro-androst-4-ene-17B-ol-3-one 17-(B-phenyl propionate) may be obtained by esterification of retro-testosterone with the acid chloride of fl-phenyl 'propionic acid in pyridine.

The introduction of alkoxy and aralkoxy groups into the 100:. methyl 9 3 steroid series may be carried out as follows:

(1) By the reaction of keto-retro-steroids with a suitable hydroxy compound (mono or poly), for example, in the presence of a catalyst, such as, for example, hydrochloric acid, pyridine-hydrochloric acid and p-toluene sulphonic acid, employing the method described by Ercoli et al. (J. Am. Chem. Soc., 82, 746 (1960)). In this manner, the cyclopentyl enolether of 17a-acetoxy-retroprogesterone may be produced. Another example is the production of the 3-enol benzyl ether of 6-dehydro-retroprogesterone by the action of benzyl alcohol and hydrochloric acid on 6-dehydro-retro-progesterone. Further, the cyclopentyl enol ether of retro-testosterone may be produced in this manner from retro-testosterone and cyclopentanol.

(2) By an acid-catalysed interchange reaction between the chosen hydroxy compound and preformed enol ether, for example by the method of Ercoli et al. (J. Am. Chem. Soc., 82, 746 (1960)). In this manner, 3-enol ethyl ether of retro-progesterone may be converted into 3-enol benzyl ether, or, 3-enol ethyl ether of retro-testosterone may be converted into 3-enol benzyl ether.

(3) By the reaction of keto-retro-steroids with trialkyl orthoformates with the use of an acid catalyst, for example an ethanolic hydrogen chloride solution, for example by the method of Ruyle et al. (J. Org. Chem., 25, 1260 (1960)). In this manner, 17a-[2'-methallyl]- retro-testosterone may be converted into 3ethoxy-17a- [2'-methallyl]-retro-androsta-3,S-diene-17 8-01, or retrotestosterone may be converted into 3-ethoxy-retro-androsta-3,5-diene-17fi-ol.

(4) By the reaction of a hydroxy-retro-steroid with an alcohol in the presence of a catalyst, such as, for example, an inorganic acid or p-toluene sulphonic acid, for example by the method of Sondheimer et al. (Tetrahedron, 5, (1959)). In this manner, 3-methoxy-retropregn-4-ene-20-one may be produced from 3-hydroxyretro-pregn-4-ene-20-one, 3-methoxy-retro-andr0st-4-ene- 17,6-01 from 3,1718-dihydroxy-retro-androst-4-ene, 3- methoxy-retro-pregn-4-ene-20-one from 3-hydroxy-retropregn-4-ene-20-one, and 3-methoxy-retro-androst-4ene- 1718-01 from 3,17B-dihydroxy-retro-androst-4-ene.

(5) By the reaction of a hydroxy-retro-steroid with diazomethane in the presence of a catalytic amount of fluoboric acid by the method of Neeman et a1. (Tetrahedron, 6, 36 (1959)). Thus 3-methoxy-retro-pregn-4-ene-20-one is obtained from retro-pregn-4ene-3-ol-20-one. Further, 3-hydroxy-retro-androst-4-ene-17-one may be converted into 3-methoxy-retro-androst-4-ene-17-one.

The production of halo-retro-steroids may be carried out as follows:

(1) By the addition of halogen to double bonds in retro-steroids, for example by addition of chlorine to 3-hydroxy-A -retro-steroids or addition of mixed halogens,

such as BrF and IF, to A -retro-steroids according to Bowers et al. (J. Am. Chem. Soc., 82, 4001 (1960)). By this method, 17u,21-dihydroxy-retro-pregna-4,9(11)-diene-3,20-dione al-acetate may be converted into the 9, ll-dihalo compounds. Thus, for example, 9(11)-dehydro-retro-testosterone may be converted into 9,11-dihaloretro-testosterone.

(2) -By the substitution-halogenation at an allyl position adjacent to a double bond or of a methylene group adjacent to a keto group in retro-steroids, for example with bromine, N-bromo-succinimide, dibromo-dimethylhydantoin etc., for example by the method of Djerassi et al. (J. Am, Chem. Soc., 72, 4534 (1950)) for the production of 6-halo-3-keto-A -steroids from 3-keto-A -steroids. Thus, retro-progesterone 20-cyclo-ethylene ketal may be brominated to form the 6-bromo compound and retro-testosterone 17-acetate may be converted into 6- bromo-retro-testosterone 17-acetate. Furthermore, retropregn-5-ene-3fi-al-20-one 3-acetate may be converted into 17a-bromo-retro-pregn-5-ene-3 8-ol-20-one 3-acetate, the double bond being protected during the reaction by conversion of the dibromide by the method of Engel et al. (Can. J. Biochem. Physiol. 35, 1047 (1957) and Can. J. Chem. 38, 452 (1960)).

(3) By the reaction of enol esters, enol ethers and enamines with halogenating agents, such as, for example, halogen, N-halo-imides, perchlorylfluoride, BrF. In this manner, 3-enol esters of 3-keto-A -retro-steroids may be converted with perchlorylfiuoride according to the method of Bloom et a1. (Chem. a.'Ind. 1959, 1317) into the 3-keto-A -6-fluoro-retro-steroids. By this method, the 3-enol acetate of retro-progesterone may be converted into 6uand 6fi-fluoro-retro-progesterone, and 3-enol acetate of retro-testosterone 17-acetate may be converted into 6aand -flu0ro-retro-testosterone 17-acetate.

Furthermore, the 3-enol ethers of 3-keto-A -retrosteroids may be converted with N-halo-imides by the method of Ringold et al. (J. Am. Chem. Soc., 81, 3485 (1959)) into the 6-halo-3-keto-A -retro-steroids. Thus, reaction of 3-ethyl enol ether of 17a-acetoxy-retro-progesterone with N-chlorosuccinimide gives 6-Cl'1lOIO-17aacetoxy-retro-progesterone. Further, reaction of 3-enol ethyl ether of retro-androst-4-ene-3,17-dione with N- chlorosuccinimide yields 6-chloro-retro-androst-4-ene-3, 17-dione.

Furthermore, the enamines of 3-keto-A -retr-o steroids may be converted by a reaction With perchlorylfiuoride followed by isomerisation with, for example, hydrochloric acid by the method of Joly et al. (Bull. 1961, 569) into 3-keto-4-fluoro A -retro-steroids. Thus, the 3-pyrrolidyl- A -enamine of 17a-acetoxy-retro-progesterone yields the 4-fiuoro-17a-acetoxy-retro-progesterone, and 3-pyrrolidyl- A -enamine of retro-testosterone yields 4-fluoro-retrotestosterone.

(4) By the reaction of retro-steroids of the following type: retro-steroid where R is an alkyl group and R may be H or 0 i J0R' where R; may be an alkyl group, which may be obtained by condensation of a methylene group activated by a keto-group with a monoor dicarboxylic acid ester (for example ethylformate, diethyloxalate), with halogenating agents, such as halogen and perchlorylfluoride, with subsequent splitting off of the carboxylic acid group. Thus, according to the method of Kissman et al. (I. Am. Chem. Soc. 82, 2316 (1960)), the sodium 23 salt 2l'ethoxallyl-retro-progesterone may be converted with perchloryldluoride and subsequent treatment with potassium acetate into 21-fiuoro-retro-progesterone. Substitution of bromine for the perchlorylfiuoride gives 21- bromo-retro-progesterone. Further, according to the method of Edwards et al. (I. Am. Chem. Soc. 81, 5262 (1960)) 2-fluoro-retro-testosterone may be produced from the sodium salt of 2-hydroxymethylene-retro-testosterone by reaction with perchlorylfiuoride and subsequent treatment with potassium acetate.

By cleavage of epoxides, for example with hydrohalogenic acids or BF for example by the method of Romo et al. (J. Org. Chem. 21, 902 (1956)), according to which 16(17)-epoxides are treated with HBr. Thus 16(17)-epoxy-retro-progesterone may be converted into 16-bromo-17a-hydroxy-retro-progesterone. According to the method of Bowers et al. (Tetrahedron 3, 14 (1958)), 6-fluoro-3,l7-diacetoxy-retro-androstane-5-ol may be produced from 3,17-diacetoxy-5(6)-epoxy-retro-androstane.

The introduction of oxo-groups in retro-steroids may be carried out as follows:

(1) By oxidation of hydroxy-retro-steroids, for example according to Oppenauer as described for the normal series by Shepherd et al. (J. Am. Chem. Soc. 77, 1212 (1955)). By this method, retro-pregn-5-ene-3-ol-20-one may be converted into retro-progesterone, or retro-androst- 5-ene-3-ol-17-one may be converted into retro-androst-4- ene-3,17-dione. The oxidation may also be performed with chromic acid, for example in pyridine, by the method of Sondheimer et al. (Tetrahedron 5, (1959) Thus, hydroxy-retro-pregn-4-ene-3-one may be converted into retro-progesterone. Further, from retro-testosterone will be obtained androst-4-ene-3,17-dione. The oxidation of hydroxy-groups occupying an allyl position with respect to a double bond, may suitably be performed with manganese dioxide, for example by the method of Sondheimer et al. (J. Am. Chem. Soc. 75, 5930 (1953)) and with quinones, such as, for example, dichloro-dicyanobenzoquinone as described by Burn et al. (Tetrahedron Letters 9, 14 (1960)). Thus, retro-pregn4-ene-3,20-diol may be converted into 20-hydroxy-retro-pregn-4-ene-3- one, and according to these two manners, retro-androst-4- ene-3,l7B-diol may be converted into retro-testosterone.

(2) By oxidation of N double bonds for example by ozonisation with the formation of 17-ketones, employing the method of Pederson et al. (J. Am. Chem. Soc. 79, 115 (1957) Thus, ozonisation of the ZZ-cyanohydrin of retro bisnorchola 4,17(20)-diene-3-on-22-al gives the retro-androst-4-ene-3,l7-dione.

(3) By hydrolysis of enol esters or enol ethers of retrosteroids, for example by acid hydrolysis as described by Serini et al. (Ber. 71, 1766 (1938)). Thus, 3-ethoxy-retropregn-3,5-diene-20-one may be hydrolysed to retro-progesterone. Further, 3-ethoxy-retro-androsta-3,S-diene-l7-one may be hydrolysed to retro-androst-4-ene-3,17-dione.

(4) By the microbiological oxidation of hydroxy-retrosteroids, for example as described by Pcrlman (Science, 115, 529 (1952)). Thus, retro-pregn-4-ene-3-ol-20-one may be converted into retro-progesterone, or 17-acetoxyretro-androst-5-ene-3-ol may be converted into retrotestosterone l7-acetate. The production of acylthio-retrosteroids may be effected by the method of Dodson et al. (J. Am. Chem. Soc. 81, 1224 (1959)), for example for the introduction of 1- and 7-acylthio groups. Thus, 6-dehydro-retro-progesterone after reaction with ethanethiolic acid gives 7-acethylthio-retro progesterone. Further, 6-dehydro-retro-testosterone after reaction with ethanethiolic acid yields the 7-acetylthio-retro-testosterone.

The production of amino-retro-steroids may be effected by reduction of retro-steroid-oximes, for example with LiAlH as described by Shoppee et al. (J. Chem. Soc., 1956, 1649). Thus, the dioxime of 6-dehydro-retro-progesterone yields 3,20-diamino-retro-pregna-4,6-diene. Further, the oxime of retro-testosterone with LiAlH, in ether yields the 3-amino-retro-androstl-ene-1713-01.

24 The production of monoalkyland dialkyl-substituted amino loot-methyl, 9B-steroids may be effected by reacting aminoor monoalkyl substituted amino 10u-methyl, 9/3.-

steroids with alkyl halides, for example with an alkyl.

bromide. Thus 3-amino-retro-androstan-173-01 when reacted with methyl bromide yields 3-(Nmethylamino)- retro-androstan-l7fl-ol.

The production of retro-steroid [2,3-d] isoxazoles may be performed by the method of Clinton et al. (J. Org. Chem. 26, 279 (1961)), by reacting 3-keto-2-hydroxymethylene retro-steroids with hydroxylamine hydrochloride. Thus, Z-hydroxymethylene-17aallyl-retro-testosterone may be converted into 17-hydroxy-17a-allyl-retroandrost-4-eno [2,3-d] isoxazole. Further, Z-hydroxymethylene-retro-testosterone may be converted into 175-hydroxy-retro-androst-4-eno [2,3-d] isoxazole.

The production of retro-steroid [3,2-c] pyrazoles may be performed by the method of Clinton et al. (J. Am. Chem. Soc. 81, 1513 (1959) by reacting 3-keto-2-hydroxymethylene retro-steroids with hydrazine. Thus, 2-hydroxymethylene-retro-pregnl-ene-20-ol-3-one by reaction with hydrazine yields ZO-hydroxy-retro-pregn-4-en-o [3,2-c] pyrazole. Further, reaction of 2-hydroxymethylene-6-dehydroretro-testosterone with hydrazine gives 17;8-hydroxyretro-androsta-4,6-dieno [3,2-c] pyrazole.

The preparation of oxido 10u-methyl-9fi steroids can be performed by reacting a double bond between the carbon atoms under consideration with an oxidizing agent such as hydrogen peroxide in the presence of an alkali or with an organic peracid such as monoperphthalic acid. Thus 6-dehydro-retro-progesterone when reacted with monoperphthalic acid yields 6,7 oxido retro-progesterone.

The introduction of methylene groups into IOa-methyl steroids may be effected by the addition of diazomethane to the double bond present in such a steroid and cleavage of the thus obtained pyrazolines according to the procedure of Wiechert and Kaspar (Chem. Ber. 93, 1810 (1960)). Thus 16-dehydro-retro-progesterone may be converted into 16-methylene-retro-progesterone.

The introduction of thiol groups (-SH) into methyl 9,3 steroids may be effected as follows:

The reaction of a steroidtosylate with thiourea and splitting of the thus formed isothiouromium compound with alkali hydroxide affords the desired thiol 10c; methyl 9;? steroid. Thus 3-thioloandrost-4-en-1713-01 17-acetate can be prepared from 3-tosyloxyandrost-4-en-1713-01 17- acetate.

The introduction of the thio analogues of alkoxy and aralkoxy groups into 100: methyl 9;? steroids can be effected by reacting an OXO-lOoc methyl, 9,8-steroid with an alkyl thiol or an aralkyl thiol respectively in the presence of a catalyst such as p-toluene sulphonic acid. Thus retrotestosterone when reacted with ethyl mercaptane yields 3-enol ethyl thio ether and when reacted with benzylmercaptane yields 3-enol benzylthioether of retro-testosterone.

The preparation of 2'-methyl-(3,2-d)-thiazoles may be carried out according to the process described by Doorenbos et al. (J. Pharm. Sc. 50, 271 (1961)) by reacting 2-bromo-3-keto retro-steroids with thiacetamide. Thus 2- bromo 17a methyl-retro-testosterone may be converted into 2' methyl (3,2-d) thiazolo 17 a methyl retro androsta-2,4-dien-17-ol.

The synthesis of 2'-amino (3,2-d) -pyrimidino retrosteroids may conveniently be performed by the reaction of 2-hydroxy methylene-3-keto-retro-steroids with guanidine according to the process described in Bull. Off. de la Trop. Ind. 1, 344 (1961). Thus 2-amino-(3,2-d)-pyrimidino-retro-androsta-4,6-dien-17fi-ol 17-acetate can be prepared from 2 hydroxymethylene-6-dehydro-retro-testosterone 17-acetate.

The production of 6-nitro retro-steroids may be effected by nitration of 3,5-dienylacylates of retro-steroids as described by Bowers et al. (I. Am. Chem. Soc. 81, 3707 25 (1959)). Thus, 3,17a-diacetoxy-retro-pregna-Zi,5-dien-20- one can be converted into 6 nitro 17a: acetoxy-retroprogesterone.

The introduction of an oxygen atom in the 18-methyl group in retro-steroids can for instance be carried out by the irradiation of a ll-nitrite of ll-hydroxy-retro-steroids according to the process described by Barton et al. (I. Am. Chem. Soc. 82, 2641 (1960)). Thus retro-corticosterone ZI-acetate can be esterified into the corresponding 11- nitrite. Irradiation of this compound in toluene yields the retro-aldosterone 2l-acetate oxime, which can be transformed by the action of sodium nitrite in acetic acid into retro-aldosterone 21-acetate.

Retro-17-spirolactones may be prepared by reaction of a 17a-acetylenic substituted retro-steroid with an alkyl Grignard halide, decomposing the complex thus formed with carbon dioxide and after hydrogenation lactonizing the thus formed hydroxy-acid, as described by Cella et al. (J. Org. Chem. 24, 743 (1959)). Thus 17a-ethinyl-3,17- dihydroxy-retro-androst ene is converted into 3,17- dihydroxy-retro-androst-S-en-17a-ylpropynoic acid, after which catalytic hydrogenation yields 3-(3,l7-dihydroxyretro-androst-S-en 17a yl) propenoic acid lactone. Repeated hydrogenation gives the corresponding 17-propanoic lactone, which by means of an Oppenauer-oxidation can be converted into the 3-keto-A -retro spirolactone.

Trihalo methyl-retro-steroids can be prepared by the reaction of cool ethers of unsaturated ketones with tetrahalo methanes as described by Lusberg et al. (Tetrahedron 9, 149 (1960)). Thus the reaction of the A -enolethyl ether of retro-progesterone with tetrabromomethane yields 6-tribromomethyl-retro-progesterone. In the same manner the trifiuoroand trichloro compounds may be prepared. The latter compound can by splitting ofi HBr and/ or reduction be converted in 6methyl-retroprogesterone.

The methods of preparing the novel retro steroids of our invention will now be explained in greater detail in the examples which follow.

Example 1 (a) By 'very vigorous stirring, 125 g. of A4,7,22-1umistatrien-3-one were dissolved at 10 C. in 2.2 litres of dry propanol-Z, which had previously been saturated with dry hydrochloric acid gas. The hydrochloric acid gas was slowly passed through the liquid for an additional half hour. The liquid was then decanted as rapidly as possible, while stirring, into a mixture of solid sodium bicarbonate and a saturated bicarbonate solution, obtained by stirring 4 kg. of sodium bicarbonate with 8 litres of water. After a short period of time the yellowish propanol-2 layer was separated out and the salt layer extracted twice with one litre of petroleum ether. The combined layers were then washed three times with a sodium bicarbonate solution and water, then dried on sodium sulphate and evaporated to dryness after filtering. The ultraviolet absorption spectrum exhibited a maximum at 287 Inn, of which the is 562 (content of pure substance about 85%).

The residue obtained was dissolved in 250 ml. of boiling petroleum ether (40 to 60 C.) and crystallized at 5 C. for a few hours and finally overnight at C. Filtering yielded 80.5 g. of lumista-4,6,22-trien-3one with a melting point of 99 to 100.5 C. (yield 64%). A portion of the last substance was recrystallized a few times with petroleum ether for analysis, the melting point rising to 101-102 C. The further analytical values were:

Found: C. 85.67%;H, 10.65%;C, 85.68%; H, 10.77%. Calculated for C H O: C, 85.22%; H, 10.73%.

In the infrared spectrum there were found apart from the 964 emfband for the side chain trans-ethylene bond three characteristic bands at 1586, 1622 and 1661 CHIS-1.

(b) While stirring vigorously, a solution of 3.0 g. of

26 lnmista-4,6,22-trien-3one in 300 ml. of dry diethylether was added to 450 ml. of liquid ammonia. During the addition of the first portion of the solution crystallization occurred but the solid dissolved after a larger portion had been added. Then, while stirring vigorously, a solution of 420 mg. of lithium in 100 ml. of ammonia was carefully added in drops until no spontaneous decoloring occurred any more. ml. of dry ethanol were then added dropwise, stirring was continued for 30 minutes, the mixture was diluted with water and the reduction product was dissolved in diethyl ether; The ethereal extracts were then extensively washed with water, dried on Na SO filtered and evaporated to dryness yielding a light-yellow resin.

( i'l.1L (A max. 242 m )=196) The resin was dissolved in ml. of boiling ethanol and after the addition of 6 ml. of 2 N NaOH the solution was boiled for five minutes. The solution was then rapidly cooled. By diluting with water, absorbing in diethyl ether, washing of the ethereal layers with water, drying, filtering and evaporating to dryness, a light-brown residue Was finally obtained with EH3, (A max. 242 m )=420 This substance was chromatographed in 25 ml. of pure benzene on 30 g. of A1203 (III) and eluated with the same solvent (total 75 ml.), a dark brown ring remaining at the upper end of the column. The dry eluate was recrystallized with 45 ml. of methanol at 5 C., after which the filtered product was washed with 20 ml. of methanol of 25 C. The yield was long light-brown needles (1.5 to 2 cms. in length), weight 2.43 g. and melting point 122 to 124 C.

500 mg. of this substance were recrystallized twice with 3 ml. of acetone at 5 C., after which finally 313 mg. of colorless crystals of lumista-4,22-dien-3-one were obtained.

The analytic values found for this pure substance were: e()\ 242 m )=16,800, melting point 122-124 C. [a] 125 (CHCI Found: C, 84.98%; H, 19.96%; C, 84.89%;H, 11.03%. Calculated for C H O: C, 84.79%; H, 11.17%.

In the infrared spectrum an intense band was found at 1665 cm." a slightly weaker band at 1620 cmf a weak band at 962 cmf and a slightly more intense hand at 978 cm.

20 g. of lumista-4,22-dien-3-one was dissolved in a mixture of 750 ml. of freshly distilled methylene chloride and 5.75 ml. of dry pyridine. The mixture, while being stirred magnetically at 80 C. (carbon dioxide ice and acetone) for 4 and /2 hours was ozonised (0.205 mol ozone per min.), speed of passing oxygen (V )=9 to 10 l./hour. The ozonide formed was then decomposed at 0 C. for one hour by stirring it with a suspension of 20 g. of zinc powder, after the addition of ml. of glacial acetic acid. The reaction mixture was then warmed for 5 minutes at 35 C., and then the resultant solid substance was separated out by filtering. After the addition of ice the filtrate was Washed successively with 75 ml. and 50 ml. of ice cold 10% Na CO solution, three times with 50 ml. of 10% NaOH solution at 0 C. and four times with 300 ml. of ice water.

The methylene chloride layer was then dried on Na SO filtered and the filtrate was distilled to dryness, the last part of which was carried out in vacuo. The colorless, practically completely crystalline residue was recrystallized with 50 ml. of diethylether at 25 C., the first crystallate being 10.1 g. of 3-ketoretrobisnorchol-4-en-22-al with a melting point of 119 to 121 C. This material was further purified by a few recrystallizations with petroleum ether (boiling region 40 to 60 C.) to which a few drops of ethyl alcohol had been added, and with diethylether.

The analytic values of this pure substance were:

M.P. 122-130 C. 6()\ max. 242.5 m )=16,700 [a] =144 C. (CHCl Found: C, 79.87%; H, 9.73%; C, 80.06%; H, 9.81%. Calculated for C H O C, 80.42%; H, 9.82%.

The infrared spectrum showed characteristic bands at 1712, 1642 and 1610 cmr (c) A solution of 450 mg. of 3keto-retro-bisnorchollen-22-al obtained as described in Example 1b in 15 ml. of chloroform and 25 ml. of acetic acid, in which 200 mg. of chromic acid and 0.2 ml. of water had been dissolved 'was oxidized at about 30 C. for 16 hours.

After the excess quantity of chromic acid had been decomposed by stirring for 30 minutes with 1.5 ml. of methanol, the mixture was diluted with water and the substance was dissolved in benzene. The benzene extract was washed with water to neutral reaction, dried on Na SO and filtered. After evaporation to dryness a crystalline residue was obtained, which by crystallization with diethylether yielded 340 mg. of 3-ketoretrobisnor-chol-4-enic acid, melting point 194 to 198 C. A few crystallizations raised the melting point of this substance to 202 to 204 C.

The further analytic values found for this pure substance were:

6()\ max.=242 m )=16,800.

Calculated for C H O C, 76.70%; H, 9.36%. Found: C, 76.93%; H, 9.45%; C, 76.92%; H, 9.48%.

(d) A solution of 450 mg. of an ozonide in 25 ml. of methylenechloride obtained in the manner described in Example 1b, by ozonizing lumista-4,22-dien-3-one, was oxidized with a solution of 200 mg. of chromic acid in 25 ml. of acetic acid overnight at 30 C. The excess quantity of chromic acid was then decomposed by stirring with 2 ml. of methanol for 30 minutes.

After dilution with water, the mixture was dissolved in diethyl ether and the ether methylene chloride extract was washed three times with 50 ml. of 2% NaOH. These layers were combined, extracted again with diethyl ether to remove residues of neutral constituents. The organic acid was freed by acidifying with concentrated hydrochloric acid and dissolving in diethylether. A-fter neutral washing with water, the mixture was dried on Na SO filtered'and finally evaporated to dryness; the yield was 240 mg. of crystalline residue. A crystallization with methanol at C. yielded 150 mg. of 3-keto-retro-bisnorchol-4-enic acid with a melting point of 200 to 203 C. When this acid was mixed with 3ketoretro-bisnorchol-4- enic acid as obtained by the method described in Example lc no drop in the melting point occurred.

(e) 0.11 ml. of'dry, freshly distilled piperidine and 1 to 5 mg. of p-toluene sulfonic acid were added to a solution of 300 mg. of 3keto-retro-bisnorchol-4-en-22-al as obtained by ozonization of lumista-4,22-dien-3-one. This solution was then refluxed in 5 ml. of dry benzene for 3 hours under N The refluxing benzene was dried by being passed through powdered BaO in an extraction column.

After cooling, the reaction mixture was poured out into water, dissolved in diethylether and then washed adequately four times with water. The ether-benzene layer, dried on Na SO and filtered, was dried to complete dryness in vacuo. The resinous residue was stirred with 3 ml. of methanol at C., cooled to C. and kept atthat temperature for 2 hours and crystallized. Filtering of this solution yielded 185 mg. of needles with a melting point of 88 to 93 C. of 22-(N-piperidyl)- retro-bisnorchola-4,20(22)dien-3-one.

Two recrystallizations with methanol raised the melting point of this substance to 94-96" C.

The analytical values of this pure substance were:

Found: C, 81.72%; H, 10.64%; N, 3.60%; C, 81.78%; H, 10.67%; N, 3.66%. Calculated for C H NO: C, 81.97%; H, 10.45%; N, 3.54%.

The infrared spectrum exhibited an intensive band at 1660 cmf which overlapped distinctly a band with lower extinction at 1650 cm. Apart from a band at 16 10 cm. a weaker band was found at 874 cmf After a few small charges of the above-mentioned pi- 28 peridino compound were produced in a similar manner, in which the final substance had a melting point of 94 to 96 0, production of several more larger charges of this compound in a different manner was carried out. The

resultant compound which had a different melting point (114 to 115 C.) was then obtained in large yields. Probably this effect is due to cis-trans isomerism.

This alternate method of preparing this compound was carried out as follows:

A solution of 10 g. of 3keto-retro-bisnorchol-4-en-22-al was refluxed in 180 ml. of dry benzene with 3.8 rnls. of piperidine and 30 mg. of p-toluenesulphonic acid for 3 hours under nitrogen. The refluxing benzene was dried by passing through powdered barium oxide.

The reaction mixture was evaporated to complete dryness in vacuo, a crystalline residue of 12.3 g. being obtained. Recrystallization with 10 ml. of methanol, yielded 9.3 g. of 22 (N-piperidyl)retro-bisnorchola-4,20(22)- dien-3-one, melting point 98 to 107-'111 C. A portion of this substance, which was sufliciently pure for further processing, was recrystallized a few times with acetone for analysis until a constant melting point of 1l4-ll5 C. was attained.

The analytical values of this substance were:

e()\ max. 241.5 m;r)=22,000, [a] =123 (CHCl C, 81.87%; H, 10.49%; N, 3.42%. C, 81.98%; H, 10.47%; N, 3.63%.

The infrared spectrum of this substance differed only on details from that of the compound having a melting point of 94 to 96 C. The characteristic bands found for said substance were also found in this case.

(f) 0.5 g. of freshly melted sodium acetate was added to a solution of 1 g. of 3keto-retro-bisnorchola-4-en-22- al, obtained as described in Example 1b, in 50 ml. of freshly distilled acetic acid anhydride. This mixture was then refluxed in a nitrogen atmosphere.

The solvent was then distilled off as far as possible under reducedpressure (about 10 minutes) and the residue obtained 'was dissolved in 25 ml. of chloroform. Undissolved sodium acetate was filtered off, the filter was washed with a small quantity of chloroform and the filtrate was diluted with the same solvent to 125 1111., a solution of 22-acetoxy-retro-bisnorchola-4,20(22)-dien'3- one being obtained. 7

(g) A solution of 300 mg. of 22-(N-piperidyl)-retrobisnorchola-4,20(22)dien-3-one in 4.5 ml. of dry thiophene-free benzene, was added dropwise at a temperature of -5 C. to l-5 C., in 45 minutes, while stirring, to a solution of 453 mg. of sodium bichromate dihydrate in 4.5 ml. of acetic acid and 3 ml. of benzene. After additional stirring, for 2 hours at 0 C., 0.75 ml. of methanol was added to the dark-colored solution and stirring was continued for 30 minutes also at 0C.

This reaction mixture was then processed by pouring it out into 25 ml. of water and by extraction twice with benzene. The combined benzene extracts were then washed successively with water, 3 ml. of cold 10% NaOH solution, twice with water, 3 ml. of cold 10% hydrochloric acid solution and four times with water. The solution was then dried on Na- SO filtered and evaporated to dryness producing a completely crystalline residue.

This residue was dissolved in 1 ml. of methylene chloride and 4 ml. of petroleum ether was added at boiling temperature. The crystallization which was completed at 25 C. yielded 150 mg. of light-yellow, hard crystal blocks of retro-progesterone with a melting point of 161v to 163 C.

A repeated recrystallization of this substanceyielded mg. with a melting point of 163164 C.

The analytical values of this pure substance were: e( \max 241.5 m L)=16,800, [a] =62 (CHCl Found: C, 79.90%; H, 9.75%. C, 79.89%; H, 9.75%. Calculated for C H O C, 80.20%; H, 9.62%.

The infrared spectrum exhibited strong bands at 1690 cm.- and 1662 cm. and a weaker band at 1615 CH1.'1.

(h) 0.5 g. of freshly melted sodium acetate was added to a solution of 1 g. of 3-keto-retro-bisnorchol-4-en-22- a1, obtained as described in Example 1b, in 50 ml. of freshly distilled acetic acid anhydride and the mixture was refluxed for 16 hours in a nitrogen atmosphere.

Then the solvent was distilled otf as far as possible under reduced pressure (about mm. Hg) and the residue obtained was dissolved in 25 ml. of chloroform.

The undissolved sodium-acetate was filtered ed, the filter was washed again with a small quantity of chloroform and the filtrate was increased with the same solvent to 150 mls. A solution of 22-acetoxy-retro-bisnorchola-4,20'- (22)-dien-3-one was obtained.

While cooling with ice water this solution was ozonized for 14 minutes, 10.5 mg. of ozone being absorbed per minute. Then, after the addition of ml. of acetic acid and 2 g. of zinc substance for 10 minutes, the mixture was shaken and after filtering the solution was washed with 10% NaOH so ution and water to neutral reaction. The solution was dried on Na SO filtered and evaporated to dryness, and then refluxed to hydrolyse any 3- enolacetate present for 45 minutes in 45 ml. of methanol and 25 ml. of 2 N H 50 After concentration in vacuo down to half the volume the solution was dissolved in diethylether. The ethereal extract was washed with 10% NaOH solution and water to neutral reaction dried on Na SO evaporated to dryness and finally crystallized with 3 ml. of diethyl ether. The crystallate obtained was recrystallized with a mixture of methylene chloride and n-hexane, the yield being 113 mg. of substance with a melting point of 160163 C. The substance did not exhibit a reduction in melting point with the retroprogesterone obtained as described in Example 1g.

[a] =-61.5 (CHCl (mean value of -62.1 and -60.9).

The following compounds listed with their starting materials are prepared according to the methods of Example 1:

7-dehydro-retro-progesterone from lumista-4,7,22-trien- 3-one;

6-dehydro-retro-progesterone from lumista-4,6,22-trien- 3-one;

1-dehydro-retro-progesterone from lumista-1,4,22-trien- 3-one;

retro-pregna-l,4,6-triene-3,20-dione from lumista-1,4,6,-

22-tetraen-3-one;

retro-pregnane-3,20-dione from lumist-22-en-3-one;

2-methyl-retro-progesterone from 2-methyl-lumista-4,6,-

22-trien-3-one;

6-methyl-retro-progesterone from 6-methyl-lumista- 4,22-dien-3-one;

6-fluoro-retro-progesterone from 6-fluorolumista-4,22-

dien-3-one;

6-chloro-retro-progesterone from 6-chloro-lumista-4,22-

dien-3-one;

6-hydroxy-retro-progesterone from 6-hydroxy-lumista- 4,22-diene-3-one;

4-chloro-retro-progesterone from 4-chloro-lumista-4,22-

dien-3-one;

1-methyl-retro-progesterone from l-methyl-lumista-r,

22-dien-3-one;

2-hydroxy retro-progesterone from Z-hydroxy-lumista- 4,22-dien-3-one;

retro-pregn-5-ene-3,20-dione from lumista-5,22-dien-3- one;

9 fiuoro-l 1-hydroxy-retro-progesterone from 9-fluoro- 1 1- hydroxy-lumista-4,22-dien-3-one;

16-hydroxy-retro-progesterone from 16-hydroxy-lumista- 4,22-dien-3-one;

l6-methyl-retro-progesterone from 16-methyl-lumist a- 4,22-dien-3-one;

30 3-hydroxy-retro-pregn-S-en-ZO-one from 3-hydroxylumista-5,22-diene and 3-hydroxy-retro-pregnan-20- one from 3-hy-droxy-lumist-22-ene.

Example 2 (a) To 3.2 ml. of anhydrous ethanol there was added a mixture of ml. of dry benzene and 12ml. of 2.86 sodium methoxide in dry methanol. This mixture was then concentrated by evaporation under N to about 30 ml. After cooling, while stirring, the resultant paste had added to it, 8.6 m1. of freshly distilled diethyloxalate, so that the reaction mixture became quite clear. Then, a solution of 10 g. of retro-progesterone, prepared as described in Example 1g, in 70 mls. of dry benzene was quickly added and stirring was continued for about minutes. By the rapid dropwise addition of 400 ml. of dry diethylether, the precipitation of the sodium enolate was completed. After stirring for an additional 45 minutes the resultant substance was filtered and washed adequately with ml. of dry diethylether. After drying, for 15 hours, on concentrated H 50 10.8 g. of yellow sodium enolate of 2l-ethoxy-oxalyl-retro-progesterone was obtained.

(b) The sodium enolate derivative thus obtained was dissolved at -20 C., while stirring, in a nitrogen atmosphere, in ml. of dry methanol. At the same temperature a solution of 5.9 g. of iodine in 210 ml. of dry methanol was added within 40 minutes to the reaction mixture while stirring and stirring was continued for 90 minutes. The decomposition of the iodine compound was carried out by adding 6.6 ml. of 3.56 N sodium-methoxide solution stirring was continued at 0 C. for one hour in dry methanol. From the solution obtained the reaction product was precipitated by adding dropwise, while stirring for 45 minutes 90 ml. of water and by then adding 120 g. of sodium chloride.

The filtered precipitate, after being washed adequately with water, was dried overnight, the yield being 5.55 g. of 21-iodo-retro-progesterone. From the filtrate, after one night, another 2.35 g. of this compound was obtained.

(c) The first fraction of 21-iodo-retro-progesterone (weight 5.55 g.) obtained this way, was refluxed in 200 ml. of purified dry acetone (purified with KMnO and K CO for 18 hours with 12 g. of freshly prepared potassium acetate purified by remelting. After evaporation of the acetone, 300 ml. of water were added and the oil separated was dissolved in methylene chloride. The extract, after drying on Na SO was evaporated to dryness (weight 1.4 g.) and chromatographed in benzene on 10 g. of neutralized A1 0 Elu-ation with benzene yielded 700 mg. of residue, from which after recrystallization with acetone, 15 0 mg. of a substance with a melting point of 159 to 164 C. was obtained.

Corresponding acetylation of the post-fraction thus obtained of 21-iodo-retro-progesterone (2.35 g.) yielded 2.6 g. of crude acetoxy-compound, from which 90 mg. of solid substance with a melting point of 160161 C. could be crystallized. Chromatography of the mother liquor as described above, after recrystallization with a mixture of acetone, and petroleum ether (40-60 C.) yielded 400 mg. with a melting point of 160-163 C.

Recrystallization of the combined crystallates with acetone yielded 425 mg. with a melting point of 164-167 C., from which, by recrystallization with ethanol, finally 335 mg. of pure retro-desoxycorticosterone acetate with a melting point of 165168 C. were obtained.

The analytical values of this pure substance were:

eE'Yi (A max=242 m 457 and 462, e 242 m =17,000 Calculated for C H O (372.49): C, 74.15%; H, 8.66%. Found: C, 74.03%; H, 8.55%; C, 74.27%; H, 8.77%.

The infrared spectrum exhibited bands at 1226, 1609, 1663, 1724 and 1751 Cm.

21-acetoxy-6-dehydro-retro-progesterone from 6-dehydro-retro-progesterone; 2l-propionoxy-l,6-bisdehydro-retro-progesterone from l,6-bisdehydro-retro-progesterone; 2l-butyroxy-retro-pregnane-3,30dione from retro-pregnane-3,ZO-dione; the hemi-succinate of 21-hydroxy-6-methyl-retroprogesterone from 6methyl-retro-progesterone; 2 1-acetoxy-9-fluoro-l l-hydroxy-retro-progesterone from 9-fiuoro-11-hydroxy-retro-progesterone; 21-formate of 21-hydroxy-2-methyl-retro-progesterone,

from 2-methyl-retro-progesterone; 2l-acetoxy-17-hydroxy-retro-progesterone from 17-hydroxy-retro-progesterone; 9-fluoro-retro-hydrocortisone 21-acetate from 9-fiuoro l 1,17-dihydroxy-retro-progesterone; 21-acetoxy-16-methyl-9-fiuoro-l 1,17-dihydroxy-retroprogesterone from 16-methyl-9-fiuoro-11,17-dihydroxy-retro-progesterone and 2 l-acetoxy-9-fluoro-1 1,16,17-trihydroxy-retroprogesterone from 9-fiuoro-11,16,17-trihydroxyretro-progesterone.

From all the above named compounds there can be prepared the unesterified corresponding 21-hydroxy derivatives by mild hydrolysis methods described in literature, for instance by reacting the esters with dilute alkali as potassium hydroxide or potassium bicarbonate.

Example 3 (a) A solution of 7.5 g. of retro-progesterone in 500 ml. of freshly distilled tertiary butyl alcohol was refluxed with 12.75 g. of finely powdered chloranil, while stirring, for hours in a nitrogen atmosphere. After cooling, 2 litres of water were added and extraction was performed three times with 200 ml. of methylene dichloride. The combined extracts were then diluted with 1 litre of petroleurn ether (4060 C.) washed successively with 100 ml. of diluted N21 SO (100 ml.) four times with 75 ml. of 1 N NaOH and water to neutral reaction.

By drying this solution on Na SO and evaporating to dryness (last part in vacuo) 3.7 g. of crystalline residue was obtained.

This residue was then dissolved in benzene.

Filtration in benzene filtered through via 35 g. of alumina (according to Brockmann, strength 11) and then the alumina was eluated with benzene. Evaporationof r the benzene yielded 3.11 g. of crystalline residue. By crystallization with 15 ml. of acetone at room temperature at lower temperatures a by-product crystallized out) 900 mg. of crystallate with a melting point of 165-170 C. were obtained. Transfer of the acetone mother liquor into a mixture of ethanol and hexane yielded 1.7 g. of asolid substance with a melting point of 130 to 145 C. This solid was then recrystallized with acetone at room temperature, yielding 600 mg. of a solid with a melting point of 166 to 171 C. The two fairly pure fractions (600 mg. and 900 mg.) yielded, after crystallization with a mixture of acetone and hexane, finally 1.0 'g. of 6-dehydroretro-progesterone, melting point 169 to 170 C. From the mother liquors an additional fraction of 0.44 g. with a melting point of 168 to 169 C. was obtained.

A small portion of the 6-dehydro-retro-progesterone was recrystallized with acetone for analysis and had the following analytical values:

Melting point 169170 C,

Elfi (A max=286.5 m )=842 and 848 e()\ 286.5 m t) 26,400.

Calculated for C H O (311.43): C, 80.73%; H, 9.03%. Found: C, 81.11%; H, 9.20%; C, 81.25%; H, 9.07%.

The infrared spectrum exhibited inter alia bands at 1695 crnf (strong, non-conjugated keto-group), 1656 cm.- (strong, conjugated keto-group), 1617 em. (strong, double bond in conjugation), 1574 em.- (moderately strong, double bond) and 888 cm. (strong).

According to the method'described in this Example 3a the following compounds listed with their starting materials are prepared:

1,6-bisdehydro-retro-progesterone from l-dehydroretro-progesterone; 2methyl-6-dehydro-retro-progesterone from Z-methylretroprogesterone; 1,6-bisdehydro-17a-hydroxy-retro-progesterone l7-acetate from 1-dehydro-17a-hydroxy-retro-progesterone 17-acetate; 6-dehydro-9-fluoro-retro-hydrocortisone from 9-fluororetro-hydrocortisone; 6-dehydro-16-hydroxy-9-fiuoro-retro-prednisolone from 16-hydroxy-9-fiuoro-retro-prednisolone; 6-dehydro-16-methyl-9-fiuoro-retro-prednisolone from 16-methyl-9-fluoro-retro-prednisolone and 6-dehydro-retro-desoxycorticosterone 21-acetate from retro-desoxycorticosterone 21-acetate.

. (b) 3.95 g. of the isolumisteronc obtained as .described in Example 1a were dissolved in 150 ml. of dry methylene dichloride. To this solution there was then added 0.81 ml. of dry pyridine. The solution thus obtained was cooled to C. and then, within 26 minutes, 15 mmol. of ozone (i.e. 140% of the theoretical volume) was passed through. The reaction mixture was then vigorously stirred, after which 15 g. of para-formaldehyde was added. Under constant stirring the temperature rose within 4 hours to about room temperature. After standing overnight the solid substance was filtered off, washed with petroleum ether and diethyl ether. The ether solution was then extracted with water, cold 1 N NaOH and water. After drying on Na SO this solutionwas evaporated and 3.08 g. of residue were obtained.

Ei'i (A max 286 m )=77O This residue was crystallized from acetone and then. recrystallized twice from acetone. The resultant pure 3-ketoretro-bisnorchola-4,6-dien-22-al had a melting point of 153155 C. The other analytical values of this substance were as follows:

The ultraviolet absorption spectrum had a maximum at 286 I'll 1..

tt...= 80; e=25,400

Calculated for C H O (326.48): C, 80.94%; H, 9.26%; O, 9.80%. Found: C, 81.07%; H, 9.16%; O, 9.94%; C, 80.87%;H, 9.05%; O, 10.00%.

The following bands in the infrared spectrum were found: 1410 emf (weak), 1654 cm.- (strong), 17-17 cm.- (strong), 1584 cm. (strong), 1621 cm.- (fairly strong).

(c) 3.5 g. of 3-keto-retro-bisnorchola-4,6-dien-22-al ob tained as described in Example 311 (crude crystalline form, melting point 146151 C.) were dissolved in 50 ml. of absolute benzene. To this solution there was then added 1.27 ml. of piperidine and 20 mg. of p-toluenesulphonic acid. This reaction mixture was refluxed for two and a half hours and the refluxing benzene dried with BaO. Then the solvent was distilled off in vacuo and 4.4. g. of residue was left This residue was crystallized from 30 ml. of methanol at 25 C. Yield: 2.1 g. of 22-(N-piperidyl)-retro-bisnorchola-4,6,20(22)- trien-3-one with a melting point of 127135 C. After two recrystallizations with acetone this substance was pure. The analytical values of this material were as follows:

Melting point (in vacuo) -136" C.

Ultraviolet absorption spectrum had a maximum at 287 m mam ssr and 57. 6:25.000 and 25,800

were added in drops, in 45 minutes, to an adequately stirred solution of 25.2 g. of sodium bichromate in 250 ml. of acetic acid+170 mls. of benzene cooled to C.

After two hours of stirring at 0.C. 42.0 ml. of methanol were added and stirring was continued for half an hour at 0 C. The reaction mixture was then poured out in water and extracted with ether. The extract was washed with water, diluted sodium hydroxide solution and water. After drying on Na SO the solvent was distilled off. Yield: 12.24 g. (92%) of residue. This residue was crystallized with acetone-hexane (6.6 g.) and then by two crystallizations with acetone the substance obtained was pure. The melting point of the 6-dehydro-retro-progesterone thus obtained was 168-169 C.

- (e) 3.95 g. of lumisterone were dissolved in 150ml. of dry methylene dichloride and to this solution there was added 0.81 ml. of pyridine. This solution was then-cooled to -70 C. and within 26 minutes, 14 mmol. of ozone was passed through. Then 15 g. of paraformaldehyde were added and, while stirring, the-temperature wasslowly raised to C. Theparaformaldehyde'was'filtered'otf, and washed with methylene dichloride. The filtrate was Washed .with water, 1 N NaOH solution and water. After drying on Na SO the organic solvent was distilled off, after which 3.11 g. of crystalline residue was left. This residue was recrystallized with a mixture of methylene dichloride and acetone. After three recrystallizations pure 3-keto-retro-bisnorchola-4,7-dien-22-al was obtained with a melting point of 196200 C and the following analytical values: I

The ultraviolet absorption spectrum had a maximum at 241 III/1.. 3 s

Calculated for C H O (326.48): C, 80.94%; H, 9.26%; O, 9.80%. Found: C, 80.15%; 'H, 9.1%; O, 10.10%; C, 80.28%; H, 9.09%; O, 10.04%.

The infrared absorption spectrum had bands, inter alia, at 1410 cm. (weak), 1660 'crnr lst-rong), 1610-cm":

(strong) and 1714 cm? (strong).

When the aldehyde thus obtained was converted in the manner described in Example 12 or. 30 into 22-(N-piperidyl)-retro-bisnorchola-4,7,20(22) tn'en 3 one, and this compound was oxidized with sodium bichromate in benzene and acetic acid in'tlie manner described 'inEx'ample 1g or 3d, 7-dehydro-retro-progesterone was obtained-By isomerisation of the 3-keto-4,7-dehydro-system of this latter compound into the 3-keto-4, 6-'dehydro-system with dry 'HCl-gas in anhydrous isopropanol; 'in the manner described in Example 1a, the 6-dehydro-retro-progesterone is obtained. 1

Example 4. Accordiug to the methods described in the Journal American Chemical Society 75 5369 (1953), 74, 5933 Melting point 214(s)217-218--C.

Calculated.for,C H O C, 76.32; H, 9.15; O, 14.53.

The infrared spectrum exhibited bands among others 3420,1699, 1653,1615, 1423, '1008and 869 cmf 34 (b) Microbiological hydroxylation of 17a,21-dihydroxy-retro-progesterone with Aspergillus orchraceus yielded retrohydrocortisone with a melting point .of -269 (dec.).

1%... (x max=242 111,1 :47

The infrared absorption spectrum showed bands inter alia at 3310, 1693, 1642, 1611, 1417, 1275, 1238, 1213, 1117, 1077, 1037, 1000, 94-9 and 870 cm.-

According to this process the following compounds are prepared: 11,17a-dihydroxy-retro-progesterone from 17 hydroxy-retro-progesterone, 1 dehydro-11,17a,2l-tr iliydroxy-retroprogesterone from 1-dehydro-17a,21 dihy- ,droxy-retroprogesterone and 6-methyl-11,17a,21-trihydroxy retro progesterone from 6-methyl-17a,21-dihydroxy-retro-progesterone.

Example 5 was distilled off, the last part being carried out in vacuo. The residue (3.49 g.) was crystallized from acetone hexane at -5 C. to yield 2.807 g. of a solid substance with a melting point of l50.5153 C. This substance was dissolved in ml. of benzene. l g. of silica gel was added to this solution after.filtr.ation, the solution was stirred and it was again evaporated to dryness. The residue (2.73

:g.) .was crystallized from acetone-hexane at 5 C(The melting point of the 1-dehydro-retro-progesterone.(2.14 g.) thus obtained was 153.5154.5 C. A small-quantity was recrystallized'to obtain a constant melting point and had the following analytical values:

7 Melting point 15 55.? 0. I c

The infrared absorption spectrum. exhibits, interalia, maxima at 1694, 1661, 1625, 1606, 1423, 1366, 1166, 889 and 814 cm.- 1

i (Xmax=242.5 mn)=496 and 502 424215 m =15,600 cH on Found: C, 80.12, 80.54; H, 8.99, 9.22. Calculated for C H O (312.45): C, 80.73; H, 9.03.

".By this process there are prepared l-dehydro-6-fluoro-retro-progesterone from .'6-fluoro-retro-progesterone; -l dehydro-6inethyl-retro-progesterone from 6-rnethyl-retro-progesterone; 1 1-dehydro-6-chloro-17 t-acetoxy-retro-progesterone from 6-chloro-17a-acetoxy-retro-progesterone;

' 1-dehydro-2',21-difiuoro-l7a-acefoxy-retro progesterone from 2,21-difiuoro-17b:acetoxy-retro-progesterone; 1-dehydro-16-methyl-17a-acetoxy-retro-progesterone a from 16-methyl-17ot-acetoxy-retro-progesterone;

'1-dehydro-l7a-bromo-retro-progesterone from 17a-bromo-retro-progesterone and 1 retro-'prednisolone from retro-hydrocortisone.

Example 6 progesterone in 100 ml. of ethanol (in total rinsed subsequently with 50 mls. of ethanol). Subsequently, the resultant mixture was refluxed under nitrogen gas for one hour and then 250 ml. of solvent was distilled off in the course of one hour. To the warm solution there were then added 300 ml. of water. While stirring occasionally the solution was cooled for ten minutes at room temperature and for 15 minutes in ice water. The resultant dioxime was filtered off, washed with water and dried at 100 C. in high vacuo. 2.85 g. of a substance with a melting point of 279-281 C. was produced.

A quantity (1 g.) of this substance was boiled with 30 ml. of ethanol and the residue (0.67 g.) was dissolved in boiling tetrahydrofurane. The resultant solution was filtered and to the filtrate there was added an equal quantity of petroleum-ether. The crystallization was achieved at 5 C. Yield 208 mg. of dioxime of 6-dehydro-retr0- progesterone with a melting point of 279282 C. (0.0) was produced. This compound had the following analytical values:

The infrared absorption spectrum exhibits maximum at 1642, 1 24, 1604. 1370. 980. 960. 889 and 878 661- Elfi (A max=278 m )=1080 and 1071 (methaual)e= Example 7 To a solution of 300 mg. of retro-preg'nane-3,20-dione in 20 ml. of anhydrous ethanol there were added 36.6 mg. of calcium chloride, Z H O and 16.3 mg. of sodium borohydride at a temperature of 20 C. The reaction mixture was then kept atthis temperature for 3 hours and then neutralised with '8 m1. of 2% acetic acid. A-fler filtration the resultant retro-pregnane-Sol-20-one was crystallized from ethanol+ether, a melting point of 167- 171 C. being obtained and had the following additional analytical values:

The infrared absorption spectrum showed inter alia bands at 3480, 1700, 1363, 1177, and 1073 cm.-

By this process the following compounds, shown with their starting materials are produced: 'l7a-acetoxy-retropregnan-3-ol-20-one from 17a-acetoxy-retro-pregnane-3, 20-dione, 21-acetoxy-retro-pregnan-3ol-20-one from 21- acetoxy retro pregnane 3,20 dione; 3,1l,17a,2ltetrohydroxy retropregnane 3 one from 11,l7a,21- tri'hydroxy retro pregnane 3,20 dione and 3,l7a,2|ltrihydroxy retro pregnan 3 one from 1711,21 dihyd-roxy retro pregnane 3,20 dione.

-267 and 7 Example 8 A solution of 0.6 g. of potassium hydroxide in 9 ml. of methanol was added to a solution of 2.826 g. of retroprogesterone in 1150 ml. of sodium-distilled dioxane. This mixture was shaken at a slight excess pressure with hydrogen in the presence of 0.3 g. of 10% palladium on carbon as a catalyst. After the absorption of 1 mol of hydrogen per molecule of the substance the mixture was filtered, the reduction product obtained being crystallized from a mixture of ether and hexane. Recrystallization yielded finally retro-pregnane-3,20-dione with a melting point of 108.5 (s) i1 -116" C. and the following additional analytical values.

79.76; 79.66; H, 9.87; 10.17. Cale, fo 21 a' (316.49); C, 79 7Q; H

The substance did not show absorption in the ultraviolet range. 7

Thus is produced 170: acetoxy retro pregnane 3,20- dione from 170: acetoxy retro progesterone; 2 1 acetoxy retro pregnane 3,20 dione from 21 acetoxyretro progesterone; 11,17l2 21 t-rihydroxy --retropregnane 3,20 dione from retro hydrocortisone or 170: acetoxy retro pregnane 1 ene 3,20 dione from 170: acetoxy 1 dehydro retr-oprogesterone and 1711,21 dihydroxy retro pregnane 3,20 dione from 17,21 dihydroxy retroprogesterone.

Example 9 (a) 5 g. of 6-dehydroretro-progesterone, 7.5 g. of chloranil and 25 .g. of powdered calcium carbonate were introduced into 170 ml. of isoamyl alcohol and the mixture was refluxed in a nitrogen'atmosphere, while stirring thoroughly, for 3 hours. After cooling, the precipitate was filtered oif and washed with methylene chloride. Water was added dropwise to the very dark filtrate and the solvent was azeotropically distilled 011 in vacuo at a temperature of less than 75 C. The resultant residue was absorbed in methylene-chloride and the solution was intensely washed with a sodium dithionite solution, then with water and with a N aqueous solution of sodium hydroxide. During the last-mentioned washing process emulsifying occurred, which could be counteracted for the major part by using a greater quantity of methylene chloride and by adding a saturated sodium chloride solution. The washing with the sodium hydroxide solution was continued until the washing layer no longer absorbed any color. After washing with water to neutral reaction, drying on sodium sulphate and filtering, the solvent was evaporated and 4.4 g. of a dark .green, semisolid substance was obtained. Crystallization from 25 mls. of di' ethyl-ether at 5 C. yielded 0.9 g. of agray substance a part of this substance from methanol raised the melting point to 150 155 C. From the ultraviolet spectra of the crystallizates and the initial mother liquor it was found that the former consisted for the major part of 6- dehydro-retro-progesterone, the latter, however, for 50% of the 1,6-bisdehydro compound.

The mother liquor was evaporated to dryness and introduced into ml. of a mixture of petroleum ether and methylene chloride |(4:1), arranged on a column of 70 g. of silica gel aud elu-ated as follows:

Fraction Solvent Volum Weight p.e.+CHiCl1 (4+1) 1.20 50 FB p.e.+ CHi0li (3+2) 1. 00 F0" p.e.+CH;Gh (3+2). 0.90 75 CHgCl; 0.30 I 75 1. 00 900 1. 40 450 0:65 440 0.50 E er 1.00 50 FK Methanol 0.40 200 Fraction FE with Egg, (286' m :694

is substantially pure 6dehydro-retro-progesterone. For fraction FF was found:

El? (226 m;r)=370, Ei'fi' (254 mg)=308 and Egg (299 m )=383 For fraction FG 380, 308 and 372 -respectively. Crystallization of the fraction FF from acetone-l-n-hexane at 5 C. yielded crystals (150 mgs.) with a melting point of 137'139 C. A mixed melting-point determination with the ll,6 bisdehydro-retro-progesterone produced in a different manner did not shown a reduction.

(b) A solution of 5 g. of 6-dehydro-rectro-progesterone and 5.1 g. of dihydro-quinone in 250 m1. ofdry, thiophene-free benzene was refluxed under a slow flow of nitrogen -for 6 hours. The initially orange-red colo red solution turned to green-red after about 1'.5 hon"fand to green after about 2.5 hours. During the last-meiitioiied period crystals of the ,dihydro-quinone began to' separate out.

37 After cooling to room temperature the reaction mixture was diluted with 250 ml. of benzene and then washed four times with 250 ml. of N sodium lye and 7 times with 250 m1. of water to a neutral reaction. Owing to the wash- 38 in 450 mls. of distilled peroxide-free dioxane at room temperature, in a nitrogen atmosphere, while stirring very vigorously for 30 seconds. After this 250ml. of water was added. After another minutes of rapid introing with sodium lye the reaction product had a light 5 duction of the fluoride gas, the fluoride was allowed to yellow color. The wash liquids were shaken with 250 ml. flow into the solution slowly for 30 minutes. The excess of benzene. The combined benzene layers were dried quantity of reagent was expelled from the solution by on sodium sulphate, filtered and the solvent was evapopassing nitrogen through it, after wh1ch 1 liter of water rated, the last part in the water-jet-pump vacuo. After was added. Extractions were then carried out twice with removal of the benzene traces, with the aid of an oil 200 ml. of methylene chloride and with 500 m1. of pepump, 3.16 g. of the crystalline residue was obtained, troleum ether. The combined extracts were washed twice whi h had with water, twice withan aqueous sodium bicarbonate 1% solution and twice with water. By drying on sodium sul- T G 404 S 299 phate, filtering and evaporating to dryness 7.33 g. of the This residue was recrystallized from acetone-hexane crystalline residue was obtained, which contained 5.52 at -55; C. Flterng egvashmgl W1th acetons-lggage percent f fl i v at .y1e ametmgpomto P 141.5 C. Stirring twice with 1 g. of silica gel each dis- (Eimm) 362) colored the light green solution of this crystallizate 'in The substance was dissolved in 300 mls. of a mixture 100 ml. of benzene. The residue obtained after the evapoof petroleum ether and benzene (1+1) and chromatoration of the benzene was crystallized from acetonegraphed on 240 gs. of neutral alumina.

Fraction Eluant Vol. Weight Am 1% Crystallization at 20 C. from Cryst.

(g-) E 1 cm. acetone-hexane 1 p.e. id. benzene: 0.5 2 1+1) 0.5 0.825 234 37s 0 57g M.P. 139-1452- 24:, 3 do 0.5 1.375 234 384 1 1 g M.P.131135- 34K 4 do 0.5 0.96 234.5 408 0.71g.M.P.133-137 1 5 do 0.5 0. 235 420 a an 1.5 0. 57 235.5 438 1.2 g. M.P.158162 58-K1 7 Ben 1mm 0. 5 O. 34 325. 5 451 8 do 1. 5 0. 40 235. 5 456 9 Benzeneethet (3+1) 2 0.26 239 205 Resin 1n Ether- 3 0. 39 240 269 Yellow crystalline residue 11 Ethanol 0.86 240 358 1 .4 g. N11. 206(S)-2l1215 (ethanol- 11-K1 BXEHG hexane at -5 C., which yielded 1.52 'g. of 1,6-bisdehydroretro-progesterone with a melting point of 141.5-142- 143.5 C.

Further processing of the mother liquors yielded a subsequent fraction of 410 mg., having a melting point of 138-l40 C.

A small quantity was recrystallized to obtain a constant melting point. The analytical values of the pure substance were as follows: A melting point of 142.5- 143-143.5 C. was found. The infrared absorption spectrum showed inter. alia maxima at 1704, 1655, 1608, 1582, 1463, 1410 (weak and broad), 887 and 768 cmF The band at 1655 cm." exhibits a distinct shoulder on the long-wave side.

1? (A max=254 mn)=305e(254 mu)=9,450 (CHgOH) it... (A 1115x 301 m,i =403e(3o1 m )=12,500(CH OH) Found: C, 81.02; 81.16; H, 8.54; 8.61. Calc. for C H O (310.44): C, 81.25; H, 8.44. By these processes there are produced G-dehydro-retro-prednisolone from 6-dehydrohydrocortisone; 6-dehydro-16-methyl-9-fluoro-prednisolone from 6- dehydro-l6-methyl-9-fluoro-hydrocortisone; 2-fiuoro-6-methyl-6-dehydro-retro-prednisolone from 2-fiuoro-6-methyl-6-dehydro-retro-hydrocortisone; 2,6-dimethyl l,6-bisdehydro-retro-progesterone from 2,6-dimethyl-6-dehydro-retwo-progesterone; 6-fluoro-1,6-bisdehydro-retro-progesterone from 6-fluoro 6-dehydro-retroaprogesterone; 1,fi-bisdehydro-17a-acetoxy-retro-progesterone from 6-dehydro-17a-acetoxy-retro-progesterone; 1,6-bisdehydro-21-fluoro-retro-progesterone from 6- dehydro-21-fluoro-retro-progesterone and 1,6- bisdehydro-6-chloro-17m-acetoxyretro-progesterone from 6-dehydro-6-chloro-17a-acetoxy-retroprogesterone Example 10 A rapid flow of perchlorlfiuoride was introduced into a solution of 7.5 g. of 3-enolacetate of retro-progesterone 0 Recrystallization of 34K from methanol yielded a mixture of blocks and scales, which were. separated as far as possible (fraction 34-BK and 34-SK respectively). The fraction 34-BK together with 2-K was recrystallized in order of succession from acetone-hexane, ethanolhexane and further twice from acetone-hexane. 185 mg. of 6a-fiuoro-retroprogesterone having a melting point of 148(s)150-151 C. were then obtained. This sample had the following analytical values:

The infrared absorption spectrum showed inter alia .maxima at 1705, 1683, 1428, 1354 and 974 cm.'

)=3s4 and 390 (methanol) 6 235 111,. =12,s00

Found: C, 76.20; 75.82; H, 8.95; 8.83. 'Calc. for

'Calc. for 0 141 0 330.45 @7632; H, 9.15; 0, 14.53

The fraction 58-K was recrystallized from acetonehexane at 5 C., 0.87 g. of 6-/3-fluoro-retro-progesterone with a melting point of 161'(s)163165 C. being thus obtained.

EiZ' (237 m =465 and 472 All the filtrates were collected andevaporated to dryness. The residue was dissolved with the fraction 34- SK (3.4 gs.) in 200 mls. of chloroform and to the solution was added 2.5% of ethanol. For two hours dry hydrochloric acid gas passed through the solution at room temperature. The acid was hen expelled for the major recrystallized substance having a constant melting point were:

Melting point l64166 C. (decomposition at -l C.).

The infrared absorption spectrum showed inter alia maxima at 1700, 1678, 1622, 1416, 1364, 1232, 1187, 1040, 945 and 875 cmf Eii (h max=237.5 mu)=4-Z9 and 642 (methanol) 6 (237.5 m )=15,300

Found: C, 75.38; H, 8.85. Calcd. for C H FO (332.46): C, 75.86; H, 8.79. [Three crystallization of the fraction ll-K from'acetone yielded finally 77 mg. of a compound having a melting pointof 219(s)220222 C. The infrared spectrum showed inter alia bands at 3470, 1682, 1666, 1616, 1420, 1364, 12033,0, 1183, 1065, 948 and 882 cm.- The infrared spectrum is identical to that of ,6-hydroxy-retroprogesterone obtained by the reaction of the 3-enolacetate of retro-progesterone with =monoperphthalic acid. The infrared spectra of the two preparations were also identical is measuring in a chloroformsolution.

E95,, (A max=242 mp.) =467 and 468 (methanol) e Found: C, 76.39; 76.39; H, 8.97; 904; O, 14.85; 14.78. Calc. for C H O (330.45): C, 76.32; H, 9.16; O, 14.53. (b) In 10 ml. of chloroform, containing 1% of ethanol there were dissolved 50 mg. of 6-u-fluoro-retro-- Crystallization from. acetone-hexane yielded crystals having a meltingpoint of 1 65,166 C. After mixing with 6-.fl-fiuoro-retro-progesterone no drop inmelting point occurred; i

By this process thus are prepared the 611 and 6,3

isomers of 6-fluoro-retro-hydrocortisonev from the 3- enolacetate of retro-hydrocortisone; 6-hydroxy-17a-acetoxy-retroaprogesterone from the 3-enolacetate of 170:- acetoxy retro progesterone; 6 fluoro 17,21 dihydroxy-retro-pregesterone, or ZI-acetate from the 3-enolacetate of 17,21 dihydroxy retro progesterone 21- acetate; 6,21 difluoro 17oz acetoxy retro progesterone from the 3 enolacetate of 21 fluoro- 17a-acetoxy retro progesterone, or 6 fluoro 17abrjomo retro progesterone from the 3-enolacetate of 17a bromo retro progesterone and 6 fluoro- 170; (2 -methallyl)-retro-testosterone from the 3- enolacetate of 170: (2 -methally) retro-testosterone.

Example 11 perature by adding dropwise, within about minutes,

a 10% solution of sodium hydroxide in water. After the addition of '13 ml. the solution still had a bright yellow color. This mixture was poured out into 150 ml. of waterv and extracted three times with 40 mLof ether. The ether extracts were washed with water to neutral reaction.

After drying on Na SO the ether was distilled off in vacuo almost to dryness, after which 3 gs. of dry potassium acetate and 50 mls. of acetone were rapidly added to the resultant concentrated solution of 21-iodo-17a- 'hydroxy-retro-progesterone. This hetergeneous mixture was then refluxed for 16 hours. After cooling the potassium salts were filtered off and washed with ace-- tone. The filtrate'was evaporated in vacuo almost to dry- ;ness, poured out into water and extracted from methyl- .ene chloride and ether; The extracts were washed with water to neutral reaction, dried on Na- SO and inspissated in vacuo. Yield 772 mg. of crude 17a, 21-dihydroxy-retro-progesterone 21-acetate,

E1 212.5 6111: 'f a I v Crystallization from acetone andv subsequently from alcoholic-hexane produced a melting point of 218(s) 238 C. (dec.); 3

E13,, 242.5 111,1 :416, 242.6 m r -16,160 Calculated for 0, 11,50 (338.51): c, 7110,11, 8.30;

0, 20.60. Found: c, 70.63; H, 3.35;"0, 19.72.

In the infrared spectrum bands were found inter alia at 3465, 1755, 1728,1653, 1612, 1419, 1231, 1098, 1085, 1061 and 1034 cm.--. I 1

Example 12 To a solution of 1.0 g. of 17a-hydroXy-6-dehydro-retroprogesterone in 15 ml. of tetrahydrofurane and 2.5 ml. of methanol there were added while'stirring at 4 C. to 0? C., 1.5 gs. of powdered. iodine. Thebrown solution obtained was discolored at the same temperature by dropwise addition within about 10 minutes, 0f 10 ml. of 1-0% sodium hydroxide solution. The reaction; mixture was poured out into 150 m1. of water and extracted from ether and methylene chloride. Theextract was washed to neutr-al reaction with water, dried on Na SO and evaporated to drynessin vacum'directiy'after'whicha mixture of 3 g.

6 dehydro-fi-methyl-l7a, 21-dihydroxy-retro progestrone of dry potassium acetate and 50 ml. of acetone was added to the resultant crude 2l-iodo-6-dehydro-17a-hydroxyretro-progesterone. This reaction mixture was refluxed for 16 hours. After cooling the salts were filtered off and washed with acetone. The filtrate was evaporated to dryness, absorbed in methylene chloride and ether and Washed to neutral reaction with water. After drying on Na SO the solvent was distilled off in'vacuo, after which 915 mg. of residue was obtained. Afterltwo fecrystallizations from a mixture of acetone and methylene chloride 6-dehydro- 17a, 2l-dihydroxy-retro-progesterone 21-acetate was obtained. Thesample had the following analytical values:

a melting point of -238.5244(s) 257 259 C. (dec.)

In the infraredspectrum bands were found inter alia at 1-067, 1046 and 1036 cmr By this process thus are produced 6-dehydro-1 1,17a',21-trihydroxy-retro-progesterone 21-' acetate from 6-dehydro-11,17-a-dihydroxy-retroprogesterone;

21-acetate from 6-dehydro-6-methyl-17a-hydroxy-retroprogesterone retro-prednisolone 21-acetate from 1.-v dehydro-l 1, l7a-dihydroxy-retro-progesterone; 2-fluoro6-dehydro-l 7a, 1 2 l-dihydroxy i'etro-progesterone 21-acetate from 2 -fluoro-6-dehydrol'7a-hydr'oxy-retroprogesterone; 3 g 16-methyl-6-dehydro-17a, 2l-dihydroxy-retroprogesterone ZI-acetate from 16-methyl6 dehydrol7a dihydroxy-retro-progesterone; A 9-fluoro-retro-hydrocortisone' -2-1-acetlat e from 9'-fluoro I 170:, 11-dihydroxy-retro-progesterone and 2-methyl-17a, 21-dihydroxy-retro-progesterone 21-propion-ate from 2- methyl-l7a=hydroxy-retro-progesterone.

Example 13 A solution of 216mg. of K CO in6 ml, of water was added to a suspension of 1.2 g. of 17a, 21-dihydroxy-retroprogesterone 21-acetatein-30 ml. of methanolat 24? C. The mixture was then stirred at 24 C. for 2 hours. After half an hour the solid substance apparently began dissolving and after one hour the whole quantity was practically dissolved. Then neutralization was'carriedout with 1 N acetic acid and the solution obtained was filtered ,oif. 

1. A 10A METHYL 9B, 8B, 13B, 14A HORMONAL STERIOD OF THE PREGNANE SERIES HAVING MORE THAN 19 CARBON ATOMS IN THE MOLECULE, A HALOGEN FREE C3 POSITION ON THE STERIOD NUCLEUS AND WHEREIN THE SUBSTITUENT ATTACHED TO THE C17 CARBON ATOM OF THE STEROID NUCLEUS IN THE B POSITION IS 